KR20030090794A - Cephem compounds - Google Patents

Abstract

The present invention provides a pharmaceutical composition comprising the compound of claim 1 in admixture with a compound of formula [I], or a pharmaceutically acceptable salt, a process for preparing a compound of formula [I], and a pharmaceutically acceptable carrier. Regarding the composition:

Where

A is lower alkylene or lower alkenylene;

R ¹ is lower alkyl, hydroxy (lower) alkyl, protected hydroxy (lower) alkyl, amino (lower) alkyl or protected amino (lower) alkyl;

R ² is hydrogen or an amino protecting group;

R ¹ and R ² join together to form lower alkylene;

R ³ and R ⁵ are independently amino or protected amino;

R ⁴ is carboxy or protected carboxy.

Description

Cephem compounds

It is an object of the present invention to provide novel cefem compounds and pharmaceutically acceptable salts thereof which are highly active against many pathogenic microorganisms.

It is another object of the present invention to provide a method for preparing the cefem compound and salts thereof.

A further object of the present invention is to provide a pharmaceutical composition comprising the cefem compound and a pharmaceutically acceptable salt thereof as the active ingredient.

It is a further object of the present invention to provide a method for treating an infectious disease caused by a pathogenic microorganism comprising administering said cefem compound to an infected human or animal.

Cefem compounds of the present invention are novel and can be represented by the formula:

Where

A is lower alkylene or lower alkenylene;

R ¹ is lower alkyl, hydroxy (lower) alkyl, protected hydroxy (lower) alkyl, amino (lower) alkyl or protected amino (lower) alkyl;

R ² is hydrogen or an amino protecting group;

R ¹ and R ² join together to form lower alkylene;

R ³ and R ⁵ are independently amino or protected amino;

R ⁴ is carboxy or protected carboxy.

As the target compound [I], the following characteristics are noted.

Namely, the target compound [I] includes the syn (Z type) isomer, the anti isomer (form E), and mixtures thereof. Neo-isomers (Z-type) refer to geometric isomers having a partial structure of the formula:

Wherein R ⁴ and R ⁵ are as defined above,

Anti isomer (form E) means another geometric isomer having a partial structure of the formula:

Wherein R ⁴ and R ⁵ are as defined above.

All of these geometric isomers and mixtures thereof are included within the scope of the present invention.

In the present specification and claims, partial structures of these geometric isomers and mixtures thereof are shown for the sake of convenience by the formula:

Wherein R ⁴ and R ⁵ are as defined above.

Another feature of note is that the pyrazolio moiety of compound [I] may exist in tautomeric form, and the tautomeric equilibrium may be represented by the following formula.

Wherein A, R ¹ , R ² and R ³ are as defined above.

While both tautomeric isomers are included within the scope of the present invention, in the present specification and claims, the target compound [I] is represented by the phenotype of the pyrazolio group of formula (A) for convenience.

The cefem compound [I] of the present invention can be prepared by the following method shown below.

Method 1

Method 2

Method 3

Method 4

Where

A, R ¹ , R ² , R ³ , R ⁴ and R ⁵ are each as defined above;

R ^3a is protected amino;

R ⁶ is protected carboxy;

Y is a leaving group;

X ⁻ is an anion;

R ^1a is protected hydroxy (lower) alkyl or protected amino (lower) alkyl;

R ^1b is hydroxy (lower) alkyl or amino (lower) alkyl.

Starting materials [II] and [VI] can be manufactured by the following method.

Method A

Method B

Where

A, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , Y and X ⁻ are each as defined above;

R ⁸ is protected amino;

R ⁹ is protected carboxy;

R ¹⁰ is protected amino.

Starting compounds [VII] and [XI] or salts thereof are prepared by the following Preparation Examples 1-7, 9-14, 16-18, 21-23, 25-45, 47-52, 54, 55, 57-61, It may be prepared by the method described in 62-66 and 68-76 or a similar method.

In the above and the following description of the specification, suitable examples and descriptions of various definitions included within the scope of the present invention are described in detail below.

The term "lower" means 1 to 6 carbon atoms, preferably 1 to 4 carbon atoms, unless otherwise indicated.

Suitable "lower alkyl" and "lower alkyl" sites, "lower alkyl", "protected hydroxy (lower) alkyl", "amino (lower) alkyl" and "protected amino (lower) alkyl" Straight or branched chain alkyl having 1 to 6 atom (s), for example methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, t-butyl, pentyl, t-pentyl and hexyl And more preferred here is C ₁ -C ₄ alkyl.

Suitable “hydroxy (lower) alkyl” is hydroxy (C ₁ -C ₆ ) alkyl such as hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl, 1-hydroxypropyl, 2-hydroxy Propyl, 3-hydroxypropyl, 4-hydroxybutyl, 5-hydroxypentyl and 6-hydroxyhexyl, more preferred here being hydroxy (C ₁ -C ₄ ) alkyl.

Suitable “amino (lower) alkyl” include amino (C ₁ -C ₆ ) alkyl such as aminomethyl, 1-aminoethyl, 2-aminoethyl, 1-aminopropyl, 2-aminopropyl, 3-aminopropyl, 4-aminobutyl, 5-aminopentyl and 6-aminohexyl, more preferred here are amino (C ₁ -C ₄ ) alkyl.

Suitable "lower alkylenes" of A include straight or branched chain alkylene having 1 to 6 carbon atoms, for example methylene, ethylene, trimethylene, tetramethylene, pentamethylene, hexamethylene and propylene More preferred in the straight chain alkylene having 1 to 3 carbon atoms.

Suitable “lower alkenylenes” of A are straight or branched chain alkenylenes having 2 to 6 carbon atoms, such as vinylene, propenylene, 1-butenylene, 2-butenylene, 1-pentenylene, 2-pentenylene, 1-hexenylene, 2-hexenylene, 3-hexenylene, more preferred here are straight chain alkenylene having 2 or 3 carbon atoms.

Suitable “lower alkylenes” formed by R ¹ and R ² include straight chain alkylene having 2 to 4 carbon atoms, such as ethylene, trimethylene and tetramethylene, more preferably 2 or 3 Linear alkylene having 2 carbon atoms.

Suitable "amino protecting groups" in "protected amino" and "protected amino (lower) alkyl" are acyl groups, substituted or unsubstituted aryl (lower) alkylidene, as described below [eg benzylidene, hydroxybenzylidene Etc.], aryl (lower) alkyl such as mono-, di- or triphenyl (lower) alkyl [eg benzyl, phenethyl, benzylhydryl, trityl and the like] and the like.

Suitable "acyl" include lower alkanoyls (eg formyl, acetyl, propionyl, hexanoyl, pivaloyl, etc.), mono (or di or tri) halo (lower) alkanoyls [eg chloroacetyl, trifluoro Acetyl, etc.], lower alkoxycarbonyl [e.g. methoxycarbonyl, ethoxycarbonyl, t-butoxycarbonyl, t-pentyloxycarbonyl, hexyloxycarbonyl, etc.], carbamoyl, aroyl [e.g. Benzoyl, toluoyl, naphthoyl, etc.], aryl (lower) alkanoyl [e.g. phenylacetyl, phenylpropionyl, etc.], aryloxycarbonyl [e.g. phenoxycarbonyl, naphthyloxycarbonyl, etc.], aryloxy (Lower) alkanoyls (eg, phenoxyacetyl, phenoxypropionyl, etc.), arylglyoxyloyls (eg, phenylglyoxyloyl, naphthylglyoxyloyl, etc.), aryl (lower) alkoxycarbonyl ( Optionally substituted with appropriate substituent (s) such as benzyloxycarbonyl, phenethyloxycarbonyl, p-nitrobenzyloxycarbonyl, and the like.

Suitable "protected hydroxy" in "protected hydroxy (lower) alkyl" include acyloxy groups, aryl (lower) alkyloxy groups, and the like. Suitable "acyl" sites in "acyloxy" include lower alkanoyls (eg formyl, acetyl, propionyl, hexanoyl, pivaloyl, etc.), mono (or di or tri) halo (lower) alkanoyls, [eg : Chloroacetyl, trifluoroacetyl, etc.], lower alkoxycarbonyl, such as methoxycarbonyl, ethoxycarbonyl, t-butoxycarbonyl, t-pentyloxycarbonyl, hexyloxycarbonyl, etc.], Carbamoyl and the like. Suitable “aryl (lower) alkyl” moieties in “aryl (lower) alkyloxy” include mono-, di- or triphenyl (lower) alkyl (eg benzyl, phenethyl, benzylhydryl, trityl, etc.) and the like. do.

Suitable “protected carboxys” include esterified carboxy groups and the like, and specific examples of ester moieties in the esterified carboxy groups include, for example, lower alkyl esters such as methyl esters, ethyl esters, propyl esters, isopropyl esters. , Butyl esters, isobutyl esters, t-butyl esters, pentyl esters, hexyl esters, 1-cyclopropylethyl esters and the like] (which may have appropriate substituent (s)), for example lower alkanoyloxy (lower) Alkyl esters such as acetoxymethyl ester, propionyloxymethyl ester, butyryloxymethyl ester, valeryloxymethyl ester, pivaloyloxymethyl ester, 1-acetoxyethyl ester, 1-propionyloxyethyl ester, 2-propionyloxyethyl ester, hexanoyloxymethyl ester, etc.], lower alkanesulfonyl (lower) alkyl; Termini, [e.g., 2-mesyl-ethyl ester, etc.] or mono (or di or tri) halo (lower) alkyl ester [e.g., 2-iodo-ethyl ester, and 2,2,2-trichloroethyl ester]; Lower alkenyl esters such as vinyl esters, allyl esters, etc .; Lower alkynyl esters such as ethynyl esters, propynyl esters, and the like; Aryl (lower) alkyl esters (which may have appropriate substituent (s)) [e.g. benzyl ester, 4-methoxybenzyl ester, 4-nitrobenzyl ester, phenethyl ester, trityl ester, benzylhydryl ester, bis (Methoxyphenyl) methyl ester, 3,4-dimethoxybenzyl ester, 4-hydroxy-3,5-di-t-butylbenzyl ester and the like]; Aryl esters (which may have appropriate substituent (s)) [eg, phenyl esters, 4-chlorophenyl esters, tolyl esters, 4-t-butylphenyl esters, xylyl esters, mesityl esters, cumenyl esters, etc.] It includes.

Suitable "leaving groups" include halogens (eg fluorine, bromine, iodine, etc.) or acyloxys such as arylsulfonyloxy [eg benzenesulfonyloxy, tosyloxy, etc.], lower alkylsulfonyloxy [eg mesyl] Oxy, etc.], lower alkanoyloxy [eg, acetyloxy, propionyloxy, etc.], and the like.

Suitable “anion” formate, acetate, trifluoroacetate, maleate, tartrate, methanesulfonate, benzenesulfonate, toluenesulfonate, chloride, bromide, iodide, sulfate, hydrogensulfate, phosphate and the like do.

Pharmaceutically acceptable suitable salts of the desired compound [I] are customary non-toxic salts, for example salts with bases or acid addition salts such as salts with inorganic bases, for example alkali metal salts (eg Sodium salts, potassium salts, etc.), alkaline earth metal salts (e.g. calcium salts, magnesium salts, etc.), ammonium salts, organic base salts, for example organic amine salts (e.g. trimethylamine salts, triethylamine salts, pyridine salts, picoline Salts, dicyclohexylamine salts, N, N'-dibenzylethylenediamine salts, and the like); Inorganic acid addition salts such as hydrochloride, hydrobromide, sulfate, hydrogensulfate, phosphate, etc .; Organic carboxylic acid or sulfuric acid addition salts such as formate, acetate, trifluoroacetate, maleate, tartrate, citrate, fumarate, methanesulfonate, benzenesulfonate, toluenesulfonate, and the like; And salts with basic or acidic amino acids such as arginine salts, aspartic acid salts, glutamic acid salts, and the like.

Preferred examples of the cefem compound of the present invention of formula [I] are as follows.

(1) R ¹ is lower alkyl, hydroxy (lower) alkyl, aryl (lower) alkyloxy (lower) alkyl, amino (lower) alkyl or acylamino (lower) alkyl;

R ² is hydrogen, aryl (lower) alkyl or acyl;

R ¹ and R ² join together to form lower alkylene;

R ³ and R ⁵ are independently amino or acylamino;

R ⁴ is carboxy or esterified carboxy A compound of formula [I], or a pharmaceutically acceptable salt thereof.

(2) R ¹ is lower alkyl, hydroxy (lower) alkyl, aryl (lower) alkyloxy (lower) alkyl, amino (lower) alkyl, (lower) alkanoylamino (lower) alkyl, or (lower) alkoxycarbo Ylamino (lower) alkyl;

R ² is hydrogen, aryl (lower) alkyl, lower alkanoyl or lower alkoxycarbonyl;

R ¹ and R ² join together to form lower alkylene;

R ³ and R ⁵ are independently amino, lower alkanoylamino or lower alkoxycarbonylamino;

R ⁴ is carboxy or lower alkoxycarbonyl, or a pharmaceutically acceptable salt thereof.

(3) R ¹ is (C ₁ -C ₆ ) alkyl, hydroxy (C ₁ -C ₆ ) alkyl, mono-, di- or triphenyl- (C ₁ -C ₆ ) alkyloxy (C ₁ -C ₆ ) Alkyl, amino (C ₁ -C ₆ ) alkyl, (C ₁ -C ₆ ) alkanoylamino (C ₁ -C ₆ ) alkyl, or (C ₁ -C ₆ ) alkoxycarbonylamino (C ₁ -C ₆ Alkyl;

R ² is hydrogen, mono-, di- or triphenyl (C ₁ -C ₆ ) alkyloxy (C ₁ -C ₆ ) alkyl, (C ₁ -C ₆ ) alkanoyl or (C ₁ -C ₆ ) alkoxycarbo Nil;

R ¹ and R ² join together to form (C ₁ -C ₆ ) alkylene;

R ³ and R ⁵ are independently amino, (C ₁ -C ₆ ) alkanoylamino or (C ₁ -C ₆ ) alkoxycarbonylamino;

R ⁴ is carboxy or (C ₁ -C ₆ ) alkoxycarbonyl, or a pharmaceutically acceptable salt thereof.

(4) R ¹ is lower alkyl, hydroxy (lower) alkyl or amino (lower) alkyl;

R ² is hydrogen;

R ¹ and R ² join together to form lower alkylene;

R ³ and R ⁵ are amino;

R ⁴ is a compound of carboxyl (2), or a pharmaceutically acceptable salt thereof.

(5) R ¹ is (C ₁ -C ₆ ) alkyl, hydroxy (C ₁ -C ₆ ) alkyl or amino (C ₁ -C ₆ ) alkyl;

R ² is hydrogen;

R ¹ and R ² join together to form (C ₁ -C ₆ ) alkylene;

R ³ and R ⁵ are amino;

R ⁴ is a compound of carboxyl (4), or a pharmaceutically acceptable salt thereof.

The preparation of the target compound of the present invention is described in detail below.

Method 1

Compound [I] or a salt thereof can be prepared by reacting a reaction derivative of Compound [II] or an amino group, or a salt thereof, with a reaction derivative of Compound [III] or a carboxy group, or a salt thereof.

Suitable reactive derivatives in the amino group of compound [II] include Schiff base imino or its tautomeric enamine type isomers obtained by reacting compound [II] with a carbonyl compound such as aldehyde, ketone or the like; Silyl derivatives obtained by reacting compound [II] with silyl compounds such as bis (trimethylsilyl) acetamide, mono (trimethylsilyl) acetamide [eg N- (trimethylsilyl) acetamide], bis (trimethylsilyl) urea, and the like ; Derivatives obtained by reacting compound [II] with phosphorus trichloride or phosgene.

Suitable salts of compound [II] and reactive derivatives thereof may be mentioned as exemplified for compound [I].

Suitable reactive derivatives in the carboxy group of compound [III] may include acid halides, acid anhydrides, activated amides, activated esters and the like. Suitable examples of these reactive derivatives include acid chlorides; Acid azide; Substituted phosphoric acids [e.g. dialkylphosphoric acid, phenyl phosphoric acid, diphenylphosphoric acid, dibenzyl phosphoric acid, halogenated phosphoric acid, etc.], dialkyl phosphorous acid, sulfurous acid, thiosulfic acid, sulfuric acid, sulfonic acid [e.g. Mixed acids with acids such as carboxylic acids (e.g. acetic acid, propionic acid, butyric acid, isobutyric acid, pivalic acid, pentanic acid, isopentanoic acid, 2-ethylbutyric acid or trichloroacetic acid, etc.) or aromatic carboxylic acids (e.g. benzoic acid, etc.) anhydride; Symmetric acid anhydrides; Activated amides with imidazole, 4-substituted imidazole, 1-hydroxy-1H-benzotriazole, dimethylpyrazole, triazole or tetrazole; Or activated esters [eg cyanomethyl ester, methoxymethyl ester, dimethyliminomethyl [(CH ₃ ) ₂ N ⁺ = CH-] ester, vinyl ester, propargyl ester, p-nitrophenyl ester, 2, 4-dinitrophenyl ester, trichlorophenyl ester, pentachlorophenyl ester, mesylphenyl ester, phenylazophenyl ester, phenylthio ester, p-nitrophenyl thioester, p-cresyl thioester, carboxymethyl thioester, pyra Nyl esters, pyridyl esters, piperidyl esters, 8-quinolyl thioesters, and the like], or N-hydroxy compounds [eg N, N-dimethylhydroxylamine, 1-hydroxy-2- (1H) -Pyridone, N-hydroxysuccinimide, N-hydroxyphthalimide, 1-hydroxy-6-chloro-1H-benzotriazole, and the like. These reactive derivatives can be arbitrarily selected from the above-mentioned derivatives according to the kind of compound [III] used.

Suitable salts of compound [III] and reactive derivatives thereof may be mentioned as exemplified for compound [II].

The reaction is generally water, alcohol (e.g. methanol, ethanol, etc.), acetone, dioxane, acetonitrile, chloroform, methylene chloride, ethylene chloride, tetrahydrofuran, ethyl acetate, N, N'-dimethylformamide, pyridine or It is carried out in other organic solvents that do not adversely affect the reaction. These conventional solvents can also be used in mixtures with water.

In the present reaction, when the compound [III] is used in free acid form or salt form thereof, the reaction is performed by conventional condensing agents such as N, N'-dicyclohexylcarbodiimide; N-cyclohexyl-N'-morpholinoethylcarbodiimide; N-cyclohexyl-N '-(4-diethylaminocyclohexyl) carbodiimide; N, N'-diethylcarbodiimide, N, N'-diisopropylcarbodiimide; N-ethyl-N '-(3-dimethylaminopropyl) carbodiimide; N, N'-carbonyl-bis- (2-methylimidazole); Pentamethyleneketene-N-cyclohexylamine; Diphenylketene-N-cyclohexylimine; Ethoxyacetylene; 1-alkoxy-1-chloroethylene; Trialkyl phosphites; Ethyl polyphosphate; Isopropyl polyphosphate; Phosphorus oxychloride (phosphoryl chloride); Phosphorus trichloride; Thionyl chloride; Oxalyl chloride; Lower alkyl haloformates (eg, ethyl chloroformate, isopropyl chloroformate, etc.); Triphenylphosphine; 2-ethyl-7-hydroxybenzisoxazolium salt; 2-ethyl-5- (m-sulfophenyl) isoxazolium hydroxide intramolecular salt; 1- (p-chlorobenzenesulfonyloxy) -6-chloro-1H-benzotriazole; It is preferably carried out in the presence of a so-called Vilsmeier reagent prepared by reacting N, N'-dimethylformamide with thionyl chloride, phosgene, trichloromethyl chloroformate, phosphorus oxychloride, and the like.

The reaction can also be carried out with inorganic or organic bases, for example alkali metal bicarbonates, tri (lower) alkylamines (eg triethylamine, diisopropylethylamine, etc.), pyridine, N- (lower) alkylmorpholine, N, N -Di (lower) alkylbenzylamine or the like.

The reaction temperature is not critical and the reaction is generally carried out under cooling or warming.

Method 2

Removal of the hydroxy protecting group is carried out on compound [Ia] or a salt thereof to prepare compound [Ib] or a salt thereof.

The removal reaction is carried out according to conventional methods such as hydrolysis, reduction and the like.

The present hydrolysis reaction is preferably carried out in the presence of an acid comprising a base or a Lewis acid.

Suitable bases include alkali metals (e.g. sodium, potassium, etc.), alkaline earth metals (e.g. magnesium, calcium, etc.), hydroxides, carbonates or bicarbonates thereof, trialkylamines (e.g. trimethylamine, triethylamine, etc.), picoline , 1,5-diazabicyclo [4.3.0] -non-5-ene, 1,4-diazabicyclo [2.2.2] octane, 1,8-diazabicyclo [5.4.0] undec-7- Yen and the like.

Suitable acids may include organic acids (eg formic acid, acetic acid, propionic acid, trichloroacetic acid, trifluoroacetic acid, etc.) and inorganic acids (eg hydrochloric acid, hydrobromic acid, sulfuric acid, hydrogen chloride, hydrogen bromide, etc.).

Removal with Lewis acids such as trihaloacetic acid (e.g. trichloroacetic acid, trifluoroacetic acid, etc.) is preferably carried out in the presence of cation trapping agents (e.g. anisole, phenol, etc.). do.

The reaction is generally carried out, for example, in water, dichloromethane, alcohols (eg methanol, ethanol, etc.), methylene chloride, tetrahydrofuran, mixtures thereof or solvents that do not adversely affect the reaction. Liquid bases or acids may also be used as solvents.

The reaction temperature is not critical and the reaction is generally carried out under cooling or warming.

Method 3- (i)

Compound [VIII] or a salt thereof can be prepared by reacting Compound [VI] or a salt thereof with Compound [VII] or a salt thereof.

Suitable salts of compounds [VI] and [VIII] may be mentioned as exemplified for compound [I].

The reaction is carried out with solvents, phosphate buffers, acetone, chloroform, acetonitrile nitrobenzene, methylene chloride, ethylene chloride, formamide, N, N-dimethylformamide, methanol, ethanol, diethyl ether, tetrahydrofuran, dimethyl sulfoxide, Or other organic solvents which do not adversely affect the reaction, preferably those having a strong polarity. Hydrophilic solvents in the solvent can be used in admixture with water. When compound [VII] is a liquid it can be used as a solvent.

The reaction is preferably carried out in the presence of a base such as an inorganic base such as an alkali metal hydroxide, an alkali metal carbonate, an alkali metal hydrogencarbonate, an organic base such as trialkylamine and the like.

The reaction temperature is not critical and the reaction is carried out at ambient temperature, cooling or heating. The invention is preferably carried out in the presence of alkali metal halides (e.g. sodium iodide, potassium iodide, etc.), alkyl metal thiocyanates (e.g. sodium thiocyanate, potassium thiocyanate, etc.).

Anion X ⁻ may be derived from leaving group X and may be converted to another anion by conventional methods.

Method 3- (ii)

Compound [I] or a salt thereof can be prepared by removing the compound [VIII] or a salt thereof by removing a carboxy protecting group.

The removal reaction is carried out in a similar manner to the reaction in Method 2 mentioned above, and the reagents and reaction conditions used (eg solvent, reaction temperature, etc.) can be referred to that of Method 2 .

Method 4

The compound [Id] or a salt thereof can be prepared by removing the compound [Ic] or a salt thereof by removing a hydroxy protecting group or an amino protecting group.

Suitable removal reaction methods include conventional ones such as hydrolysis, reduction and the like.

(i) hydrolysis:

The present hydrolysis reaction is preferably carried out in the presence of an acid comprising a base or a Lewis acid.

Suitable bases include alkali metals (e.g. sodium, potassium, etc.), alkaline earth metals (e.g. magnesium, calcium, etc.), hydroxides, carbonates or bicarbonates thereof, trialkylamines (e.g. trimethylamine, triethylamine, etc.), picoline , 1,5-diazabicyclo [4.3.0] -non-5-ene, 1,4-diazabicyclo [2.2.2] octane, 1,8-diazabicyclo [5.4.0] undec-7- Yen and the like.

Suitable acids may include organic acids (eg formic acid, acetic acid, propionic acid, trichloroacetic acid, trifluoroacetic acid, etc.) and inorganic acids (eg hydrochloric acid, hydrobromic acid, sulfuric acid, hydrogen chloride, hydrogen bromide, etc.).

Removal with Lewis acids such as trihaloacetic acid (eg trichloroacetic acid, trifluoroacetic acid, etc.) is preferably carried out in the presence of cation trapping agents (eg anisole, phenol, etc.). do.

The reaction is generally carried out, for example, in water, dichloromethane, alcohols (eg methanol, ethanol, etc.), methylene chloride, tetrahydrofuran, mixtures thereof or solvents that do not adversely affect the reaction. Liquid bases or acids may also be used as solvents.

The reaction temperature is not critical and the reaction is generally carried out under cooling or warming.

(ii) reduction:

Reduction is carried out in conventional manner, including chemical reduction and catalytic reduction.

Suitable reducing agents for use in chemical reduction include metals (e.g. tin, zinc, iron, etc.) or metal compounds (e.g. chromium chloride, chromium acetate, etc.) and organic or inorganic acids (e.g. formic acid, acetic acid, propionic acid, trifluoroacetic acid). , p-toluenesulfonic acid, hydrochloric acid, hydrobromic acid, etc.).

Catalysts suitable for use in catalytic reduction include platinum catalysts (e.g. platinum plates, sponge platinum, platinum black, colloidal platinum, platinum oxides, platinum wires, etc.), palladium catalysts (e.g. sponge palladium, palladium black , Palladium oxide, palladium on carbon, colloidal palladium, barium sulfate palladium, barium carbonate palladium and the like], nickel catalyst [e.g., reduced nickel, nickel oxide, Raney nickel, etc.], cobalt catalyst [e.g., reduced cobalt, Raney Cobalt, etc.], iron catalysts (eg, reduced iron, Raney iron, etc.), copper catalysts (eg, reduced copper, Rani copper, Ullman copper, etc.).

The reduction is generally carried out in conventional solvents which do not adversely affect the reaction, for example water, methanol, ethanol, propanol, dioxane, tetrahydrofuran, N, N-dimethylformamide or mixtures thereof.

In addition, when the above-mentioned acids used for chemical reduction are liquids, they may also be used as solvents.

In addition, suitable solvents used in catalytic reduction may be the solvents mentioned above, and other conventional solvents such as diethyl ether, dioxane, tetrahydrofuran, or the like, or mixtures thereof.

The reaction temperature of this reduction is not critical and the reaction is generally carried out under cooling or warming.

When R ⁵ is protected amino The amino protecting group in R ⁵ can be removed by conventional methods, for example by hydrolysis.

Methods A and B for the preparation of starting compounds are described in detail below.

Method A- (i)

Compound [XII] or a salt thereof can be prepared by reacting Compound [X] or a salt thereof with Compound [XI] or a salt thereof.

The removal reaction is carried out in a manner similar to the reaction in the above-mentioned method 3- (i), and the reagents and reaction conditions (eg, solvent, reaction temperature, etc.) used may refer to those of the method 3- (i). .

Method A- (ii)

Compound [II] or a salt thereof can be prepared by removing reaction of the amino protecting group of R ⁸ and R ¹⁰ and the carboxy protecting group of R ⁹ with compound [XII] or a salt thereof.

The removal reaction is carried out in a manner similar to the reaction in Method 2 mentioned above, and the reagents used and the reaction conditions (eg solvent, reaction temperature, etc.) can be referred to that of Method 2 .

Method B

Compound [VI] or a salt thereof can be prepared by reacting a reaction derivative in compound [XIII] or an amino group, or a salt thereof with a reaction derivative in compound [XIV] or a carboxy group, or a salt thereof.

The removal reaction is carried out in a similar manner to the reaction in Method 1 mentioned above, and the reagents and reaction conditions used (eg solvent, reaction temperature, etc.) can be referred to that of Method 1 .

The compound obtained by the above method can be separated or purified by conventional methods such as milling, recrystallization, column chromatography, recrystallization and the like.

Compound [I] and other compounds may comprise one or more stereoisomer (s) such as optical isomer (s) and geometric isomer (s) due to asymmetric carbon atom (s) and double bond (s) and All such isomers and mixtures thereof are included within the scope of the present invention.

Compounds of interest [I] and pharmaceutically acceptable salts thereof include solvates [e.g., including compounds (e.g. hydrates, etc.)].

The desired compound [I] and its pharmaceutically acceptable salts are novel and exhibit high antimicrobial activity by inhibiting the proliferation of various pathogenic microorganisms, including Gram-positive and Gram-negative microorganisms, and thus antimicrobial agents, in particular antioral agents. It is useful as a microbial agent.

In order to demonstrate the usefulness of the desired compound [I], test data for urine excretion and MIC (minimum inhibitory concentration) of each representative compound of the present invention are shown below.

Test Methods :

In vitro antibacterial activity is measured by the following double agar plate dilution method.

One loop of each test strain (10 ⁶ viable cells per ml) incubated overnight in tryticase-soy broth was lined on a cardiac leaching agar (HI-agar) containing a gradient of test compound, 37 ° C. After 20 hours of incubation, the minimum inhibitory concentration is expressed in μg / ml.

Test compound:

Compound (a): 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxyimino) acetami Fig.]-3- [3-amino-4- (3-aminopropyl) -2-methyl-1-pyrazolo] methyl-3-cepem-4-carboxylate

Compound (b): 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxyimino) acetami Fig.]-3- [3-amino-4- (2-aminoethyl) -2-methyl-1-pyrazolio] methyl-3-cepem-4-carboxylate

Test result:

Table 1

Test strain Test compound MIC (μg / ml) Pseudomonas aeruginosa FP 2056 (a) One (b) 0.5

For therapeutic purposes, the desired compound [I] and pharmaceutically acceptable salts thereof of the present invention are in admixture with a pharmaceutically acceptable carrier such as an organic or inorganic solid or liquid excipient suitable for oral, parenteral and external administration, As active ingredients the compounds are used in the form of conventional pharmaceutical preparations. Pharmaceutical formulations may be prepared in solid form such as tablets, granules, powders, capsules, or in the form of solutions such as solutions, suspensions, syrups, emulsions, lemonades and the like.

If necessary, the preparations may contain auxiliaries, stabilizers, wetting agents and lactose, citric acid, tartaric acid, stearic acid, magnesium stearate, terra alba, sucrose, corn starch, talc, gelatin, agar, pectin, peanut oil, olive oil. And other commonly used additives such as cacao butter, ethylene glycol and the like.

The dosage of the compound of formula [I] may vary depending on the age, condition, type of disease, and type of compound [I] applied to the patient. Generally, an amount of from 1 mg to about 4,000 mg per day can be administered to the patient. An average single dose of 50 mg, 100 mg, 250 mg, 500 mg, 1000 mg of the target compound [I] of the present invention can be used to treat diseases infected with pathogenic microorganisms.

Through the following Preparation Examples and Examples will be described in more detail the present invention.

The present invention relates to novel cefem compounds and pharmaceutically acceptable salts thereof. More particularly, the present invention relates to novel cefem compounds having antimicrobial activity and pharmaceutically acceptable salts thereof, methods for their preparation, pharmaceutical compositions comprising them, and methods of treating infectious diseases in humans and animals.

Preparation Example 1

Triphenylmethyl chloride (61.3 g) in a suspension of 5-amino-1-methylpyrazole-4-carbaaldehyde (25 g, 200 mmol) and triethylamine (22.2 g, 220 mmol) in dichloromethane (500 ml) at room temperature , 220 mmol) was added. The mixture was stirred at rt for 70 h. The reaction mixture was washed successively with 10% aqueous citric acid solution, brine and 10% aqueous sodium hydrogen carbonate solution. The extract was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with ethyl acetate to give 1-methyl-5-triphenylmethylaminopyrazole-4-carbaaldehyde (67.6 g) as a colorless solid.

Preparation Example 2

Triethyl phosphonoacetate (26.9 g, 120 mmol) was added dropwise to a suspension of sodium hydride (60% dispersion in mineral oil, 4.8 g, 120 mmol) in tetrahydrofuran (200 ml) under ice cooling. The mixture was stirred for 1 h under ice cooling. 1-methyl-5-triphenylmethylaminopyrazole-4-carbaaldehyde (36.7 g, 100 mmol) was added to the reaction mixture at room temperature, and the mixture was stirred at room temperature for 17 hours. After evaporation of the solvent in vacuo, the residue was dissolved in chloroform. The solution was washed with 10% aqueous citric acid solution, brine, and 10% aqueous sodium carbonate solution. The extract was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with ethyl acetate and dried in vacuo to give ethyl (E) -3- (1-methyl-5-triphenylmethylaminopyrazol-4-yl) acrylate (33.0 g) as a colorless solid.

Preparation Example 3

Ethyl (E) -3- (1-methyl-5-triphenylmethylaminopyrazol-4-yl) acrylate in a suspension of lithium aluminum hydride (5.7 g, 150 mmol) in tetrahydrofuran (200 ml) at room temperature (21.9 g, 50 mmol) was added. The mixture was stirred at rt for 1 h. After cooling in an ice bath potassium fluoride (34 g) and water (10 ml) were added to the reaction mixture. Insoluble matter was removed by filtration. The filtrate was concentrated in vacuo and the residue dissolved in chloroform. The solution was washed with 10% citric acid aqueous solution. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The oily residue was purified by column chromatography on silica gel eluting with chloroform to give 3- (1-methyl-5-triphenylmethylaminopyrazol-4-yl) propanol (14.8 g) as a solid.

Preparation Example 4

3- (1-methyl-5-triphenylmethylaminopyrazol-4-yl) propanol (4.0 g, 10 mmol), phthalimide (1.5 g, 10 mmol) and triphenyl in tetrahydrofuran (20 ml) Diisopropyl azodicarboxylate (3.0 g, 15 mmol) was added to the phosphine (4.0 g, 15 mmol) solution, and the mixture was stirred at room temperature for 30 minutes. To the reaction mixture was added a solution of 4 mol / l hydrogen chloride in dioxane (10 ml) and the mixture was stirred at room temperature for 30 minutes. Water was added to the reaction mixture and the mixture was washed with ethyl acetate. The organic layer was separated and the aqueous layer was adjusted to pH 9 with 10% aqueous sodium carbonate solution. The solution was extracted with chloroform. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with ethyl acetate and dried in vacuo to afford 5-amino-1-methyl-4- (3-phthalimidopropyl) pyrazole (2.2 g) as a solid.

Preparation Example 5

5-Amino-1-methyl-4- (3-phthalimidopropyl) pyrazole (1.42 g, 5 mmol) was added to concentrated hydrochloric acid (10 ml) at room temperature and the mixture was stirred at reflux for 23 h. The reaction mixture was added to water (50 ml) and filtered to remove most of the insoluble matter. The filtrate was washed with ethyl acetate and the aqueous layer was concentrated in vacuo. The residue was triturated with 2-propanol and dried in vacuo to give 3- (5-amino-1-methylpyrazol-4-yl) propylamine dihydrochloride (700 mg) as a solid.

Preparation Example 6

In a solution of 3- (5-amino-1-methylpyrazol-4-yl) propylamine dihydrochloride (9.3 g, 40 mmol) in methanol (80 ml) 28% sodium methoxide in methanol (15.9 g) \ Was added. The mixture was filtered through celide and the filtrate was concentrated in vacuo. The solution of oily residue in ethyl formate (60 g) was stirred at reflux for 20 hours. After cooling, chloroform was added to the reaction mixture. The mixture was filtered through celide and the filtrate was concentrated in vacuo. The residue was triturated with ethyl acetate and dried in vacuo to give N- [3- (5-amino-1-methylpyrazol-4-yl) propyl] formamide (6.7 g) as a solid.

Preparation Example 7

A solution of N- [3- (5-amino-1-methylpyrazol-4-yl) propyl] formamide (3.64 g, 20 mmol) and triethylamine (2.23 g, 22 mmol) in methylene chloride (50 ml) To this was added triphenylmethyl chloride (6.13 g, 22 mmol). The mixture was stirred at rt for 90 min. The reaction mixture was washed with 10% aqueous citric acid solution, brine, and 10% aqueous sodium carbonate solution. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with ethyl acetate and dried in vacuo to give N- [3- (1-methyl-5-triphenylmethylaminopyrazol-4-yl) propyl] formamide (6.8 g) as a solid.

Preparation Example 8

To a solution of benzylhydryl 7β-t-butoxycarbonylamino-3-chloromethyl-3-cepem-4-carboxylate (1.03 g, 2.00 mmol) in N, N-dimethylformamide (2.5 ml), sodium iodide ( 300 mg, 2.00 mmol) was added and the mixture was stirred at rt for 30 min. To the reaction mixture was added N- [3- (1-methyl-5-triphenylmethylaminopyrazol-4-yl) propyl] formamide (2.55 g, 6.00 mmol) and methylene chloride (5 ml). The whole mixture was stirred at rt for 18 h and poured into a mixture of ethyl acetate and water. The aqueous layer was separated and the organic layer was washed with brine, dried over sodium sulfate and filtered. The filtrate was concentrated in vacuo to about 20 ml. The concentrate was poured into diisopropyl ether (300 ml) and the resulting precipitate was collected by filtration and dried in vacuo. To the resulting solid solution in methylene chloride (7.5 ml) was added anisole (2.5 ml) and trifluoroacetic acid (5.0 ml) under ice cooling. The resulting solution was stirred at rt for 3 h and poured into diisopropyl ether (300 ml). The resulting precipitate was collected by filtration and dried in vacuo to afford crude 7β-amino-3- [3-amino-4- (3-formamidopropyl) -2-methyl-1-pyrazolo] methyl-3-cepem- 4-carboxylate bistrifluoroacetic acid salt (1.67 g) was obtained. This product was used in the next step without further purification.

Example 1

Crude 7β-amino-3- [3-amino-4- (3-formamidopropyl) -2-methyl-1 in a mixed solvent of N, N-dimethylformamide (16 ml) and tetrahydrofuran (16 ml) (Z) -2- under ice cooling in a solution of -pyrazolo] methyl-3-cepem-4-carboxylate bistrifluoroacetic acid salt (1.65 g) and N-trimethylsilylacetamide (2.94 g, 22.4 mmol) (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-t-butoxycarbonyl-1-methylethoxyimino) acetyl chloride hydrochloride salt (863 mg, 2.24 mmol ) Was added. The solution was stirred for 2.5 h under ice cooling. The reaction mixture was poured into ethyl acetate (400 ml) and the mixture was stirred for 30 minutes. The resulting precipitate was collected by filtration, washed successively with ethyl acetate and diisopropyl ether, and dried in vacuo to give a solid (1.07 g).

To a suspension of the resulting solid in methylene chloride (3.0 ml) was added anisole (1.0 ml) and trifluoroacetic acid (2.0 ml) under ice cooling. The resulting solution was stirred at rt for 2.5 h and poured into diisopropyl ether (150 ml). The resulting precipitate was collected by filtration and dried in vacuo to afford the crude product, which was purified by preparative high performance liquid chromatography (HPLC) using an ODS column. The eluate containing the desired product was concentrated to about 30 ml in vacuo. The concentrate was adjusted to ca. pH 3 with concentrated hydrochloric acid and chromatographed on microporous nonionic adsorptive resin Daion ^R HP-20 (Mitsubishi Chemical Corporation) eluting with 30% aqueous 2-propanol. The eluate was concentrated to about 30 ml in vacuo and 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxy Mino) acetamido] -3- [3-amino-4- (3-formamidopropyl) -2-methyl-1-pyrazolo] methyl-3-cefe-4-carboxylate (118 mg) Obtained as an intangible solid.

Example 2

7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxyimino in methanol (1.1 ml) at room temperature Acetamido] -3- [3-amino-4- (3-formamidopropyl) -2-methyl-1-pyrazolo] methyl-3-cepem-4-carboxylate (112 mg, 0.172 mmol ) Hydrogenated hydrochloric acid (0.11 ml) was added. The mixture was stirred at rt for 4 h. The reaction mixture was adjusted to pH 6 with saturated aqueous sodium hydrogen carbonate and concentrated in vacuo to remove methanol. The resulting residue was purified by preparative HPLC using an ODS column. The eluate containing the desired product was concentrated to about 30 ml in vacuo. The concentrate was adjusted to ca. pH 3 with concentrated hydrochloric acid and chromatographed on Daion ^R HP-20 (Mitsubishi Chemical Corporation) eluting with 30% aqueous 2-propanol. The eluate was concentrated to about 30 ml in vacuo and lyophilized to afford 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1- Methylethoxyimino) -acetamido] -3- [3-amino-4- (3-aminopropyl) -2-methyl-1-pyrazolo] methyl-3-cepem-4-carboxylate (20 mg ) Obtained as an intangible solid.

Preparation Example 9

A solution of ethyl (E) -3- (1-methyl-5-triphenylmethylaminopyrazol-4-yl) acrylate (60 g, 137 mmol) in ethanol (700 ml) was heated for 10 hours at room temperature under hydrogen atmosphere. Treated with% palladium carbon (6.0 g). After filtration of the catalyst the filtrate was concentrated in vacuo. The residue was triturated with hexane in vacuo and dried in vacuo to give ethyl 3- (1-methyl-5-triphenylmethylaminopyrazol-4-yl) propionate (63 g) as a colorless solid.

Preparation Example 10

To a solution of ethyl 3- (1-methyl-5-triphenylmethylaminopyrazol-4-yl) propionate (22 g, 50 mmol) in methanol (60 ml) was added 10% aqueous sodium hydroxide solution (60 ml), The mixture was stirred at reflux for 2 hours. The reaction mixture was acidified with 10% aqueous citric acid solution and the mixture was extracted with a mixed solvent of chloroform and methanol. The extract was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with diisopropyl ether and dried in vacuo to afford 3- (1-methyl-5-triphenylmethylaminopyrazol-4-yl) propionic acid (16 g) as a colorless solid.

Preparation Example 11

Ethyl in a solution of 3- (1-methyl-5-triphenylmethylaminopyrazol-4-yl) propionic acid (14.4 g, 35 mmol) and triethylamine (4.9 ml, 35 mmol) in tetrahydrofuran (150 ml) It was added to chloroformate (3.4 ml, 35 mmol) and stirred for 20 minutes under ice cooling. To the reaction mixture was added a solution of sodium azide (2.3 g, 35 mmol) in water (30 ml) under ice-cooling. The mixture was stirred for 20 minutes under ice cooling and then for 20 minutes at room temperature. Cold water was added to the reaction mixture and the mixture was extracted with ethyl acetate. The extract was washed with saturated aqueous sodium hydrogen carbonate solution, dried over anhydrous magnesium sulfate and filtered. Methanol was added to the filtrate and the mixed solution was concentrated in vacuo. Methanol (150 ml) was added to the residue and the mixture was stirred at reflux for 1.5 h. The reaction mixture was concentrated in vacuo to afford methyl N- [2- (1-methyl-5-triphenylmethylaminopyrazol-4-yl) ethyl] carbamate (15.6 g) as a colorless solid.

Preparation Example 12

A mixture of methyl N- [2- (1-methyl-5-triphenylmethylaminopyrazol-4-yl) ethyl] carbamate (15.3 g, 34.7 mmol) and concentrated hydrochloric acid (80 ml) was refluxed for 13 hours. Stirred. The reaction mixture was washed with ethyl acetate. The organic layer was separated and the aqueous layer was concentrated in vacuo. The residue was triturated with diisopropyl ether and dried in vacuo to afford 2- (5-amino-1-methylpyrazol-4-yl) ethylamine dihydrochloride (6.1 g) as a colorless solid.

Preparation Example 13

To a solution of 2- (5-amino-1-methylpyrazol-4-yl) ethylamine dihydrochloride (2.98 g, 14 mmol) in methanol (50 ml) 28% sodium methoxide in methanol (5.4 ml, 28 mmol) Seed solution was added. The mixture was filtered through celide and the filtrate was concentrated in vacuo. The mixture of residue and ethyl formate (80 ml) was stirred at reflux for 16 h. Chloroform was added to the reaction mixture, and the mixture was filtered through celite. The filtrate was concentrated in vacuo to give N- [2- (5-amino-1-methylpyrazol-4-yl) ethyl] formamide (2.7 g) as a brown oil.

Preparation Example 14

A solution of N- [2- (5-amino-1-methylpyrazol-4-yl) ethyl] formamide (2.7 g, 16 mmol) and triethylamine (2.5 ml, 17.6 mmol) in methylene chloride (50 ml) Triphenylmethyl chloride (5.2 g, 17.6 mmol) was added to the mixture, and the mixture was stirred at room temperature for 3 hours. The reaction mixture was washed successively with 10% aqueous citric acid solution and brine. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with diisopropyl ether and dried in vacuo to give N- [2- (1-methyl-5-triphenylmethylaminopyrazol-4-yl) ethyl] formamide (4.7 g) as a colorless solid. It was.

Preparation Example 15

7β-amino-3- [3-amino-4- (2-formamidoethyl) -2-methyl-1-pyrazolo] methyl-3-cepem-4-carboxylate bistrifluoroacetic acid salt

The title compound was prepared in the same manner as in Production Example 8, benzylhydryl 7β-t-butoxycarbonylamino-3-chloromethyl-3-cepem-4-carboxylate and N- [2- (1-methyl-5- Triphenylmethylaminopyrazol-4-yl) ethyl] formamide was obtained.

Example 3

7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxyimino) acetamido] -3- [3-amino-4- (2-formamidoethyl) -2-methyl-1-pyrazolo] methyl-3-cepem-4-carboxylate

The title compound was prepared in the same manner as in Example 1, 7β-amino-3- [3-amino-4- (2-formamidoethyl) -2-methyl-1-pyrazolo] methyl-3-cepem-4 -Carboxylate bistrifluoroacetic acid salt and (Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-t-butoxycarbonyl-1- Methylethoxyimino) acetyl chloride hydrochloride salt was obtained.

Example 4

7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxyimino) acetamido] -3- [3-amino-4- (2-aminoethyl) -2-methyl-1-pyrazolo] methyl-3-cepem-4-carboxylate

The title compound was prepared in the same manner as in Example 2, 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methyl Ethoxyimino) -acetamido] -3- [3-amino-4- (2-formamidoethyl) -2-methyl-1-pyrazolo] methyl-3-cepem-4-carboxylate It was.

Preparation Example 16

To a suspension of 5-amino-1- (2-hydroxyethyl) pyrazole-4-carbonitrile (10 g, 65.7 mmol) and pyridine (100 ml) at room temperature is added triphenylmethyl chloride (22.3 g, 78.9 mmol). It was. After stirring at 60 ° C. for 6 hours, the reaction mixture was concentrated in vacuo. Tetrahydrofuran and brine were added to the residue, and the separated organic layer was washed with brine. The extract was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with ethyl acetate to give 5-amino-1- [2- (trityloxy) ethyl] pyrazole-4-carbonitrile (25.4 g) as a colorless solid.

Preparation Example 17

In a suspension of lithium aluminum hydride (4.89 g, 129 mmol) in tetrahydrofuran (318 ml) at room temperature, 5-amino-1- [2- (trityloxy) ethyl] pyrazole-4-carbonitrile (25.4 g, 64.4 mmol) was added. The mixture was stirred at reflux for 12 h. After cooling in an ice bath sodium fluoride (25.4 g), tetrahydrofuran (100 ml), dichloromethane (200 ml) and water (10 ml) were added to the reaction mixture. Insoluble matter was removed by filtration. The filtrate was concentrated in vacuo to afford {5-amino-1- [2- (trityloxy) ethyl] pyrazol-4-yl} methylamine (19.8 g) as a solid.

Preparation Example 18

A suspension of {5-amino-1- [2- (trityloxy) ethyl] pyrazol-4-yl} methylamine (19.7 g, 49.4 mmol) in ethyl formate (500 ml) was stirred at 50 ° C. for 3 hours. It was. After evaporation of the solvent in vacuo, the residue was triturated with diisopropyl ether to give N- {5-amino-1- [2- (trityloxy) ethyl] pyrazol-4-yl} methylformamide (7.99 g ) Was obtained as a solid.

Preparation Example 19

(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) in a mixture of tetrahydrofuran (80 ml) and N, N-dimethylformamide (20 ml) [(2 A solution of sodium bis (trimethylsilyl) amide (8.33 g) in tetrahydrofuran (12 ml) was added to a solution of -t-butoxy-1,1-dimethyl-2-oxoethoxy) imino] ethanoic acid (5 g). Was added and the mixture was stirred for 15 minutes. To the reaction mixture under ice cooling was added a solution of di-t-butyl dicarbonate (3.3 g) in tetrahydrofuran (20 ml) and the mixture was stirred for 3 hours under ice cooling. Ethyl acetate was added to the reaction mixture, and the mixture was washed with 10% aqueous potassium hydrogen sulfate solution and then with phosphate buffer (pH 6.86). The organic layer was separated and dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with diisopropyl ether and dried in vacuo (Z) -2- {5-[(t-butoxycarbonyl) amino] -1,2,4-thiadiazol-3-yl} [ (2-t-butoxy-1,1-dimethyl-2-oxoethoxy) imino] ethanoic acid (3.10 g) was obtained.

Preparation Example 20

A mixture of N, N-dimethylformamide (0.648 ml) and phosphoryl chloride (0.781 ml) was stirred at room temperature for 30 minutes. Tetrahydrofuran (4 ml) and (Z) -2- {5-[(t-butoxycarbonyl) amino] -1,2,4-thiadiazol-3-yl} [(4 2-t-butoxy-1,1-dimethyl-2-oxoethoxy) imino] ethanoic acid (3 g) was added and the reaction mixture was stirred at room temperature for 1 hour. At the same time, a mixture of benzylhydryl 7β-amino-3-chloromethyl-3-cepem-4-carboxylate hydrochloride (3 g) and N-trimethylsilylacetamide (8.72 g) in tetrahydrofuran (15 ml) was heated to A clear solution was prepared. The solution was then cooled to -20 ° C and added to the activated acidic solution obtained above. The reaction mixture was stirred for 1 h at a temperature of -10 ° C to 0 ° C and poured into a mixture of ethyl acetate and water. The aqueous layer was separated and the organic layer was washed with brine, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated in vacuo and benzylhydryl 7β-[(Z) -2- (5-t-butoxycarbonylamino-1, by column chromatography on silica gel eluted with hexanes / ethyl acetate (3: 2). 2,4-thiadiazol-3-yl) -2- (1-t-butoxycarbonyl-1-methylethoxyimino) acetamido] -3-chloromethyl-3-cepem-4-carboxylate (4.79 g) was obtained.

Example 5

Trimethylsilyl iodide (1.38 ml) in a solution of 5-amino-4-formamidomethyl-1- (2-triphenylmethyloxyethyl) pyrazole (2.06 g) in dichloromethane (10 ml) under ice cooling and Diisopropylethylamine (1.68 ml) was added and the mixture was stirred for 2 hours under ice cooling. Benzylhydryl 7β-[(Z) -2- (5-t-butoxycarbonylamino-1,2, in N, N-dimethylformamide (4 ml) which was stirred for 0.5 h under ice cooling to the reaction mixture. 4-thiadiazol-3-yl) -2- (1-t-butoxycarbonyl-1-methylethoxyimino) acetamido] -3-chloromethyl-3-cepem-4-carboxylate (2 g ) And sodium iodide (364 mg) mixture were added. The reaction mixture was stirred at rt for 22 h and added to a mixture of ethyl acetate and water. The organic layer was separated and dried over magnesium sulfate. Magnesium sulfate was filtered and the filtrate was concentrated to about 20 ml in vacuo. The concentrate was poured into diisopropyl ether (150 ml) and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the resulting solid in methylene chloride (9 ml) was added anisole (3 ml) and trifluoroacetic acid (6 ml). The resulting solution was stirred at rt for 3 h and poured into diisopropyl ether (300 ml). The resulting precipitate was collected by filtration and dried in vacuo. The resulting powder was dissolved in phosphate buffer (pH 7) and the solution was adjusted to about pH 6 using saturated aqueous sodium hydrogen carbonate solution. The solution containing the desired compound was purified by preparative HPLC using an ODS column. The eluate containing the desired product was concentrated in vacuo. The concentrate was adjusted to about pH 2 with concentrated hydrochloric acid and eluted with 20% aqueous 2-propanol and chromatographed on Diaion ^R HP-20 (Mitsubishi Chemical Corporation). The eluate was evaporated in vacuo and lyophilized to yield 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxy. Mino) -acetamido] -3- [3-amino-4-formamidomethyl-2- (2-hydroxyethyl) -1-pyrazolo] methyl-3-cepem-4-carboxylate (103 mg) was obtained as an intangible solid.

Example 6

7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxyimino in methanol (1 ml) at room temperature Acetamido] -3- [3-amino-4-formamidomethyl-2- (2-hydroxyethyl) -1-pyrazolo] methyl-3-cepem-4-carboxylate (97 mg) Concentrated hydrochloric acid (0.1 ml) was added to the solution. The mixture was stirred at rt for 4.5 h and poured into ethyl acetate. The resulting precipitate was collected by filtration and dried in vacuo. The resulting powder was dissolved in phosphate buffer (pH 7) and the solution was adjusted to about pH 6 using saturated aqueous sodium hydrogen carbonate solution. The solution containing the desired compound was purified by preparative HPLC using an ODS column. The eluate containing the desired product was concentrated in vacuo. The concentrate was adjusted to about pH 2 with concentrated hydrochloric acid and eluted with 20% aqueous 2-propanol and chromatographed on Diaion ^R HP-20 (Mitsubishi Chemical Corporation). The eluate was evaporated in vacuo and lyophilized to yield 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxy. Mino) acetamido] -3- [3-amino-4-aminomethyl-2- (2-hydroxyethyl) -1-pyrazolo] methyl-3-cepem-4-carboxylate (17.5 mg) Obtained.

Preparation Example 21

To the sodium hydride suspension (55% dispersion in mineral oil, 26.2 g, 600 mmol) in tetrahydrofuran (1000 ml) under ice cooling was added diethyl (cyanomethyl) phosphate (106.3 g, 600 mmol). The mixture was stirred for 20 minutes under ice cooling. 1-methyl-5-triphenylmethylaminopyrazole-4-carbaaldehyde (197 g, 530 mmol) was added to the mixture, and the mixture was stirred for 2 hours. The reaction mixture was poured into ice cold water. The mixture was extracted with ethyl acetate. The extract was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with diethyl ether to give 3- (1-methyl-5-triphenylmethylaminopyrazol-4-yl) acrylonitrile (E, Z mixture, 126 g) as a solid.

E type

¹ H-NMR (CDCl ₃ ) δ3.14 (3H, s), 4.45 (1H, br), 5.09 (1H, d, J = 16.5 Hz), 6.43 (1H, d, J = 16.5 Hz), 7.17- 7.32 (15 H, m), 7.42 (1 H, s)

Z type

¹ H-NMR (CDCl ₃ ) δ3.09 (3H, s), 4.37 (1H, br), 4.58 (1H, d, J = 11.9 Hz), 6.22 (1H, d, J = 11.9 Hz), 7.17- 7.32 (15H, m), 8.14 (1H, s)

Preparation Example 22

Toluene (1.0 mol) in a solution of 3- (1-methyl-5-triphenylmethylaminopyrazol-4-yl) acrylonitrile (E, Z mixture, 26 g, 66.6 mmol) in methylene chloride (250 ml) under ice cooling solution of diisobutylaluminum hydride in / l, 200 ml, 200 mmol) was added dropwise. The mixture was stirred at rt for 24 h. Methanol (100 ml) was added dropwise to the reaction mixture. To the mixture was added sodium fluoride (5 g) and methanol (300 ml) dropwise. Insoluble matter was removed by filtration. After evaporation of the solvent in vacuo, the oily residue was dissolved in methylene chloride. To the solution was added saturated aqueous sodium hydrogen carbonate solution (25 ml) and di-t-butyl dicarbonate (20 g). The mixture was stirred at rt for 17 h. The aqueous layer was separated and the organic layer was washed with water. The organic layer was dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography, eluting with ethyl acetate / hexanes on silica gel. The eluate was concentrated in vacuo and the residue was recrystallized from diethyl ether to give t-butyl N- [3- (1-methyl-5-triphenylmethylaminopyrazol-4-yl) -2-propenyl] carbamate (E, Z mixture, 1.8 g) was obtained as colorless crystals.

E type

¹ H-NMR (CDCl ₃ ) δ 1.47 (9H, s), 2.90 (3H, s), 3.49-3.52 (2H, m), 4.17 (1H, s), 4.18 (1H, br), 5.55-5.63 (2H, m), 7.18-7.30 (15H, m), 7.38 (1H, s)

Z type

¹ H-NMR (CDCl ₃ ) δ 1.44 (9H, s), 2.95 (3H, s), 3.76-3.80 (2H, m), 4.35 (1H, br), 4.38 (1H, br), 5.05-5.09 ( 1H, m), 5.56 (1H, d, J = 11.5 Hz), 7.18-7.29 (16H, m)

Example 7

Benzylhydryl 7β-[(Z) -2- (5-t-butoxycarbonylamino-1,2,4-thiadiazol-3-yl)-in N, N-dimethylformamide (2.1 ml)- Sodium iodide (203) in 2- (1-t-butoxycarbonyl-1-methylethoxyimino) acetamido] -3-chloromethyl-3-cepem-4-carboxylate (1.02 g, 1.20 mmol) solution mg, 1.35 mmol) was added and the mixture was stirred at rt for 30 min. To the resulting reaction mixture mixture t-butyl N- [3- (1-methyl-5-triphenylmethylaminopyrazol-4-yl) -2-propenyl] carbamate (E, Z mixture, 1.40 g, 2.80). mmol) and methylene chloride (4.2 ml) were added. The whole mixture was stirred at rt for 24 h. Ethyl acetate (100 ml) and water (50 ml) were added to the reaction mixture. The aqueous layer was separated and the organic layer was washed with brine, dried over sodium sulfate and filtered. The filtrate was concentrated to about 5 ml in vacuo. The concentrate was poured into diisopropyl ether (160 ml) and the resulting precipitate was collected by filtration and dried in vacuo. Anisole (1.43 ml) and trifluoroacetic acid (2.86 ml) were added to the resulting solid solution in methylene chloride (4.29 ml) under ice cooling. The resulting solution was stirred at room temperature for 3 hours and poured into diisopropyl ether (120 ml). The resulting precipitate was collected by filtration and dried in vacuo to give a solid (1.32 g). Purification by preparative HPLC using a solid ODS column. The first eluate containing the desired product was concentrated to about 30 ml in vacuo. The concentrate was adjusted to about pH 3 with concentrated hydrochloric acid and eluted with 30% aqueous 2-propanol and chromatographed on Diaion ^R HP-20 (Mitsubishi Chemical Corporation). The eluate was concentrated to about 30 ml in vacuo and lyophilized to afford 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1). -Methylethoxyimino) acetamido] -3- [3-amino-4-((E) -3-amino-1-propenyl) -2-methyl-1-pyrazolo] methyl-3-cepem 4-carboxylate (35 mg) was obtained as an intangible solid.

The second eluate was treated in the same manner as above to give 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methyl Toxyimino) -acetamido] -3- [3-amino-4-((Z) -3-amino-1-propenyl) -2-methyl-1-pyrazolo] methyl-3-cepem-4 -Carboxylate (14 mg) was obtained as an intangible solid.

Preparation Example 23

To a solution of 5-amino-4-aminomethyl-1-methylpyrazole dihydrochloride (10 g) in methanol (100 ml) was added a solution of 28% sodium methoxide in methanol (19.4 ml). The mixture was filtered and the filtrate was concentrated in vacuo. Separately acetic anhydride (14.2 ml) was added to formic acid (11.5 ml) and the mixture was stirred at room temperature for 30 minutes. The resulting mixture was the oily residue (5-amino-4-aminomethyl-1-methylpyrazole) obtained above, and the mixture was stirred at room temperature for 2 hours. After evaporating the solvent in vacuo, water was added to the residue. The mixture was adjusted to pH 9.5 using Diaion ^R HP-20 (Mitsubishi Chemical Corporation). The mixture was filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with dichloromethane / methanol (3: 1) to afford 5-formamido-4-formamidomethyl-1-methylpyrazole (3.97 g).

Preparation Example 24

Sodium iodide (1.02 g) in a solution of benzylhydryl 7β-t-butoxycarbonylamino-3-chloromethyl-3-cepem-4-carboxylate (3.5 g) in N, N-dimethylformamide (4.5 ml) Was added and the mixture was stirred at rt for 30 min. 5-formamido-4-formamidomethyl-1-methylpyrazole (3.71 g) was added to the reaction mixture. The whole mixture was stirred at rt for 29 h and poured into a mixture of ethyl acetate and water. The aqueous layer was separated and the organic layer was washed with brine, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated to about 20 ml in vacuo. The concentrate was poured into diisopropyl ether (300 ml) and the resulting precipitate was collected by filtration and dried in vacuo. To the resulting solid solution in methylene chloride (10.5 ml) was added anisole (3.5 ml) and trifluoroacetic acid (7 ml). The resulting solution was stirred at room temperature for 3 hours and poured into diisopropyl ether. The resulting precipitate was collected by filtration and dried in vacuo to afford crude 7β-amino-3- (3-formamido-4-formamidomethyl-2-methyl-1-pyrazolo) methyl-3-cepem-4- Carboxylate bistrifluoroacetate (3.07 g) was added. This product was used in the step without further purification.

Example 8

Crude 7β-amino-3- (3-formamido-4-formamidomethyl-2-methyl in a mixed solvent of N, N-dimethylformamide (15 ml) and tetrahydrofuran (15 ml) under ice cooling -1-pyrazolo) methyl-3-cepem-4-carboxylate bistrifluoroacetate (3.07 g) and N-trimethylsilylacetamide (6.47 g) in solvent (Z) -2- (5-amino- 1,2,4-thiadiazol-3-yl) -2- (1-t-butoxycarbonyl-1-methylethoxyimino) acetyl chloride hydrochloride salt (1.9 g) was added. The solution was stirred for 2 hours under ice cooling. The reaction mixture was added to ethyl acetate (300 ml) and the mixture was stirred for 30 minutes. The resulting precipitate was collected by filtration, washed successively with ethyl acetate and diisopropyl ether, and dried in vacuo to give a solid (3.28 g).

Anisole (3 ml) and trifluoroacetic acid (6 ml) were added to the resulting solid suspension in methylene chloride (9 ml) under ice cooling. The resulting solution was stirred at room temperature for 3 hours and poured into diisopropyl ether. The resulting precipitate was collected by filtration and dried in vacuo to afford the crude product 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1 -Methylethoxyimino) acetamido] -3- (3-formamido-4-formamidomethyl-2-methyl-1-pyrazolo) methyl-3-cepem-4-carboxylate (3.44 g ) Was obtained. This product was used for the next step without further purification.

Example 9

7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxyimino in methanol (25 ml) at room temperature ) Acetamido] -3- (3-formamido-4-formamidomethyl-2-methyl-1-pyrazolo) methyl-3-cepem-4-carboxylate (2.5 g) (2.5 ml) was added. The mixture was stirred at rt for 17 h. The reaction mixture was adjusted to about pH 7 with saturated aqueous sodium hydrogen carbonate solution and concentrated in vacuo to remove methanol. The resulting residue was purified by preparative HPLC using an ODS column. The eluate containing the desired product was concentrated to about 30 ml in vacuo. The concentrate was adjusted to pH using concentrated hydrochloric acid and eluted with 20% aqueous 2-propanol and chromatographed on Diaion ^R HP-20 (Mitsubishi Chemical Corporation). The eluate was evaporated in vacuo and lyophilized to yield 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxy. Mino) acetamido] -3- (3-amino-4-aminomethyl-2-methyl-1-pyrazio) methyl-3-cepem-4-carboxylate (50 mg) was obtained as an amorphous solid.

Preparation Example 25

To a suspension of lithium aluminum hydride (3.01 g) in tetrahydrofuran (150 ml) was added 1-ethyl-5- (tritylamino) -1H-pyrazole-4-carbonitrile (7.5 g). The mixture was stirred for 55 hours. After cooling in an ice bath sodium fluoride (13.3 g) and water (6 ml) were added to the reaction mixture. Insoluble matter was removed by filtration. The filtrate was evaporated and 4-aminomethyl-1-ethyl-5-tritylaminopyrazole (3.4 g) was added.

Preparation Example 26

To a solution of 4-aminomethyl-1-ethyl-5-tritylaminopyrazole (3 g) in tetrahydrofuran (30 ml) was added di-t-butyl dicarbonate (2.05 g). The mixture was stirred at rt for 4 h. The reaction mixture was added to ethyl acetate and water. The aqueous layer was separated and the organic layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated in vacuo and purified by column chromatography on silica gel eluting with ethyl acetate / hexanes (2: 3). The eluate was concentrated in vacuo to afford 4-t-butoxycarbonylaminomethyl-1-ethyl-5-tritylaminopyrazole (2.55 g).

Example 10

Benzylhydryl 7β-[(Z) -2- (5-t-butoxycarbonylamino-1,2,4-thiadiazol-3-yl)-in N, N-dimethylformamide (3 ml)- Of a mixture of 2- (1-t-butoxycarbonyl-1-methylethoxyimino) acetamido] -3-chloromethyl-3-cepem-4-carboxylate (1 g) and sodium iodide (199 mg) 4-t-butoxycarbonylaminomethyl-1-ethyl-5-tritylaminopyrazole (1.17 g) was added to the suspension, and the mixture was stirred at room temperature for 47 hours. The reaction mixture was added to a mixture of ethyl acetate (30 ml) and water (20 ml). The organic layer was separated, washed with brine (15 ml) and dried over anhydrous magnesium sulfate. Magnesium sulfate was filtered and the filtrate was concentrated to about 5 ml under reduced pressure. The concentrate was poured into diisopropyl ether (150 ml) and the resulting precipitate was collected by filtration and dried in vacuo. To the resulting solid solution in methylene chloride (4.5 ml) was added anisole (1.5 ml) and trifluoroacetic acid (3 ml). The resulting solution was stirred at room temperature for 3 hours and poured into diisopropyl ether. The resulting precipitate was collected by filtration and dried in vacuo. The resulting powder was dissolved in phosphate buffer (pH 7) and adjusted to about pH 6 using saturated aqueous sodium hydrogen carbonate solution. The solution containing the desired compound was purified by preparative HPLC using an ODS column. The eluate containing the desired product was concentrated in vacuo. The concentrate was adjusted to about pH 2 with concentrated hydrochloric acid and eluted with 20% aqueous 2-propanol and chromatographed on Diaion ^R HP-20 (Mitsubishi Chemical Corporation). The eluate was evaporated in vacuo and lyophilized to yield 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxy. Mino) acetamido] -3- (3-amino-4-aminomethyl-2-ethyl-1-pyrazolio) methyl-3-cepem-4-carboxylate (155 mg) was obtained as an amorphous solid.

Preparation Example 27

Triphenylmethyl chloride (23.2 g) was added to a solution of 5-amino-1-isopropyl-1H-pyrazole-4-carbonitrile (10.4 g) in pyridine (200 ml) at room temperature. The mixture was stirred at 60 ° C. for 5 hours. The reaction mixture was evaporated under reduced pressure and extracted with ethyl acetate. The extract was dried over anhydrous magnesium sulfate, filtered and evaporated in vacuo. The residue was triturated with diisopropyl ether and 1-isopropyl-5-tritylamino-1H-pyrazole-4-carbonitrile (19.1 g) was added.

Preparation Example 28

To a suspension of lithium aluminum hydride (3.09 g) in tetrahydrofuran (160 ml) was added 1-isopropyl-5-tritylamino-1H-pyrazole-4-carbonitrile (8 g). The mixture was refluxed for 72 hours. After cooling in an ice bath sodium fluoride (13.7 g) and water (6 ml) were added to the reaction mixture. Insoluble material was filtered off and the filtrate was evaporated in vacuo. To the residue solution in tetrahydrofuran (80 ml) was added di-t-butyl dicarbonate (6.68 g) and the mixture was stirred at rt for 3 h. The reaction mixture was evaporated in vacuo and the residue was purified by column chromatography on silica gel eluting with ethyl acetate / hexanes (1: 3). The eluate was concentrated in vacuo to afford 4-t-butoxycarbonylaminomethyl-1-isopropyl-5-tritylaminopyrazole (3.12 g).

Example 11

7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxyimino) acetamido] -3- (3-amino-4-aminomethyl-2-isopropyl-1-pyrazolo] methyl-3-cepem-4-carboxylate

Benzylhydryl 7β-[(Z) -2- (5-t-butoxycarbonylamino-1,2,4-thiadiazol-3-yl) -2 in the same manner as in Example 10 -(1-t-butoxycarbonyl-1-methylethoxyimino) acetamido] -3-chloromethyl-3-cepem-4-carboxylate and 4-t-butoxycarbonylaminomethyl-1- Obtained from isopropyl-5-tritylaminopyrazole.

Preparation Example 29

Triphenylmethyl chloride (36.35 g) was added to a suspension of 5-amino-1-propyl-1H-pyrazole-4-carbonitrile (16.32 g) in pyridine (160 ml) at room temperature. After stirring at 60 ° C. for 5 hours, the reaction mixture was evaporated in vacuo. The residue was added to a mixture of tetrahydrofuran and brine and the organic layer was separated and washed with brine. The extract was dried over anhydrous magnesium sulfate, filtered and evaporated in vacuo. The residue was triturated with diisopropyl ether to give 1-propyl-5-tritylamino-1H-pyrazole-4-carbonitrile (37.21 g) as a colorless solid.

Preparation Example 30

To a suspension of lithium aluminum hydride (8.94 g) in tetrahydrofuran (600 ml) was added 1-propyl-5-tritylamino-1H-pyrazole-4-carbonitrile (37 g). The mixture was refluxed for 7 days. After cooling in an ice bath, sodium fluoride (40 g) and water (17 ml) were added to the reaction mixture. Insoluble matter was removed by filtration. Ethyl formate was added to the filtrate, and the mixture was refluxed for 24 hours. After evaporation of the solvent in vacuo, the residue was triturated with diisopropyl ether and dried in vacuo to afford 4-aminomethyl-1-propyl-5-tritylaminopyrazole (30.3 g).

Preparation Example 31

To a solution of 4-aminomethyl-1-propyl-5-tritylaminopyrazole (9.6 g) in a mixture of ethyl acetate (150 ml) and methanol (30 ml) is added 4N hydrochloric acid in ethyl acetate (24.2 ml) and the mixture Was stirred at room temperature for 2 hours. The resulting precipitate was collected by filtration and dried in vacuo to afford 4-aminomethyl-1-propyl-5-aminopyrazole dihydrochloride (3.3 g).

Preparation Example 32

A solution of 4-aminomethyl-1-propyl-5-aminopyrazole dihydrochloride (3.6 g) in a mixture of tetrahydrofuran (35 ml) and water (35 ml) was adjusted to pH 9 with saturated aqueous sodium carbonate solution and To the solution was added a solution of di-t-butyl dicarbonate (6.92 g) in tetrahydrofuran (15 ml). The mixture was stirred for 1 hour while maintaining the pH of the solution with aqueous sodium carbonate solution. The reaction mixture was added to ethyl acetate and water. The aqueous layer was separated and the organic layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated in vacuo and the residue was triturated with diisopropyl ether and dried in vacuo to afford 4-t-butoxycarbonylaminomethyl-1-propyl-5-aminopyrazole (2.21 g).

Preparation Example 33

4-t-butoxycarbonylaminomethyl-1-propyl-5-tritylaminopyrazole

In the same manner as in Preparation 27, the title compound was obtained from 4-t-butoxycarbonylaminomethyl-1-propyl-5-aminopyrazole.

Example 12

7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxyimino) acetamido] -3- (3-amino-4-aminomethyl-2-propyl-1-pyrazolo] methyl-3-cepem-4-carboxylate

Benzylhydryl 7β-[(Z) -2- (5-t-butoxycarbonylamino-1,2,4-thiadiazol-3-yl) -2 in the same manner as in Example 10 -(1-t-butoxycarbonyl-1-methylethoxyimino) acetamido] -3-chloromethyl-3-cepem-4-carboxylate and 4-t-butoxycarbonylaminomethyl-1- Obtained from propyl-5-tritylaminopyrazole.

Preparation Example 34

Hydrochloric acid (1.8 ml) was added to a solution of 5-amino-1- (2-hydroxyethyl) -1H-pyrazole-4-carbonitrile (15 g) in methanol (300 ml) under ice cooling. The resulting solution was stirred for 15 minutes. The mixture was treated with 10% palladium carbon (7.5 g) under hydrogen atmosphere at room temperature for 3 hours. The catalyst was filtered off and the filtrate was concentrated in vacuo to afford 5-amino-1- (2-hydroxyethyl) -1H-pyrazole-4-carbaaldehyde (13.3 g).

Preparation 35

Triphenylmethyl chloride (86.7 g) was added to a solution of 5-amino-1- (2-hydroxyethyl) -1H-pyrazole-4-carbaaldehyde (40.2 g) in pyridine (400 ml) at room temperature. The mixture was stirred at 60 ° C. for 5 hours. The reaction mixture was evaporated under reduced pressure and extracted with ethyl acetate. The extract was dried over anhydrous magnesium sulfate, filtered and evaporated in vacuo. The residue was purified by column chromatography on silica gel eluting with ethyl acetate / hexanes (3: 2) to give 5-amino-1- (2-triphenylmethyloxyethyl) -1H-pyrazole-4-carbaaldehyde ( 62.8 g) was obtained.

Preparation Example 36

A solution of 5-amino-1- (2-triphenylmethyloxyethyl) -1H-pyrazole-4-carbaaldehyde (15 g) and ammonium acetate (4.07 g) in nitromethane (300 ml) was refluxed for 6.5 hours. The reaction mixture was evaporated under reduced pressure and extracted with dichloromethane. The extract was dried over anhydrous magnesium sulfate, filtered and evaporated in vacuo to afford 5-amino-4-[(E) -2-nitroethenyl] -1- (2-triphenylmethyloxyethyl) -1H-pyrazole (4.46 g) was obtained.

Preparation Example 37

To a suspension of lithium aluminum hydride (3.22 g) in tetrahydrofuran (100 ml) under ice cooling, 5-amino-4-[(E) -2-nitroethenyl] -1- (2-triphenylmethyloxyethyl)- 1 H-pyrazole (4.4 g) was added. The mixture was refluxed for 5 hours. After cooling in an ice bath, sodium fluoride (14.3 g) and water (6.12 ml) were added to the reaction mixture. Insoluble matter was removed by filtration. The filtrate was concentrated in vacuo to afford 5-amino-4- (2-aminoethyl) -1- (2-triphenylmethyloxyethyl) -1H-pyrazole (3.63 g). This product was used for the next step without further purification.

Preparation Example 38

Di-t-butyl dicarbonate in a solution of 5-amino-4- (2-aminoethyl) -1- (2-triphenylmethyloxyethyl) -1H-pyrazole (3.6 g) in tetrahydrofuran (36 ml) (2.02 g) was added. The mixture was stirred at rt for 19 h and evaporated in vacuo. The residue was purified by column chromatography on silica gel eluting with ethyl acetate / hexanes (2: 3). The eluate was concentrated in vacuo to afford 5-amino-4- (2-t-butoxycarbonylaminoethyl) -1- (2-triphenylmethyloxyethyl) -1H-pyrazole (1.79 g).

Example 13

Solution of 5-amino-4- (2-t-butoxycarbonylaminoethyl) -1- (2-triphenylmethyloxyethyl) -1H-pyrazole (1.24 g) in dichloromethane (5 ml) under ice cooling To trimethylsilyl iodide (0.688 ml) and diisopropylethylamine (0.842 ml) were added and the mixture was stirred for 2 hours under ice cooling. Benzylhydryl 7β-[(Z) -2- (5-t-butoxycarbonylamino in N, N-dimethylformamide (2 ml) stirred for 30 minutes under ice cooling and 18 hours at room temperature in the reaction mixture. -1,2,4-thiadiazol-3-yl) -2- (1-t-butoxycarbonyl-1-methylethoxyimino) acetamido] -3-chloromethyl-3-cepem-4 A mixture of carboxylate (1 g) and sodium iodide (182 mg) was added. The reaction mixture was added to ethyl acetate and water mixture. The organic layer was separated and dried over magnesium sulfate. Magnesium sulfate was filtered and the filtrate was concentrated to about 20 ml under reduced pressure. The concentrate was poured into diisopropyl ether (150 ml) and the resulting precipitate was collected by filtration and dried in vacuo. To the resulting solid solution in methylene chloride (3 ml) was added anisole (1 ml) and trifluoroacetic acid (2 ml). The resulting solution was stirred at room temperature for 3 hours and poured into diisopropyl ether. The resulting precipitate was collected by filtration and dried in vacuo. The resulting powder was dissolved in phosphate buffer (pH 7) and adjusted to pH 6 using aqueous sodium bicarbonate solution. The solution containing the desired compound was purified by preparative HPLC using an ODS column. The eluate containing the desired product was concentrated in vacuo. The concentrate was adjusted to about pH 2 with concentrated hydrochloric acid and eluted with 20% aqueous 2-propanol and chromatographed on Diaion ^R HP-20 (Mitsubishi Chemical Corporation). The eluate was evaporated in vacuo and lyophilized to yield 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxy. Mino) acetamido] -3- [3-amino-4- (2-aminoethyl) -2- (2-hydroxyethyl) -1-pyrazolo] methyl-3-cepem-4-carboxylate ( 66.4 mg) was obtained as an intangible solid.

Preparation Example 39

To the sodium hydride suspension (60% dispersion in mineral oil, 3.46 g) in tetrahydrofuran (250 ml) under ice cooling was added dropwise diethyl (cyanomethyl) phosphate (23.3 ml). The mixture was stirred for 1 h under ice cooling. To the reaction mixture was added 1- (2-triphenylmethyloxyethyl) -5-aminopyrazole-4-carbaaldehyde (47.76 g) in tetrahydrofuran (250 ml) at room temperature. The mixture was stirred at rt for 5 h and extracted with ethyl acetate. The extract was washed with aqueous sodium hydrogen carbonate solution, water and brine. The extract was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with diisopropyl ether and dried in vacuo to afford 4-[(E) -2-cyanoethenyl] -1- [2- (trityloxy) ethyl] -5-aminopyrazole (59.38 g) Obtained.

Preparation Example 40

To a suspension of lithium aluminum hydride (5.41 g) in tetrahydrofuran (200 ml) under ice cooling in 4-[(E) -2-cyanoethenyl] -1- [2- (trityl) in tetrahydrofuran (100 ml) A solution of oxy) ethyl] -5-aminopyrazole (15 g) was added. The mixture was refluxed for 3.5 h. After cooling in an ice bath, sodium fluoride (24 g) and water (10.3 ml) were added to the reaction mixture. Insoluble matter was removed by filtration. The filtrate was concentrated in vacuo to afford 4- (3-aminopropyl) -1- (2-triphenylmethyloxyethyl) -5-aminopyrazole (13.7 g). This product was used in the next step without further purification.

Preparation Example 41

Di-t-butyl dicarbonate (7.72 g) in a solution of 4- (3-aminopropyl) -1- (2-triphenylmethyloxyethyl) -5-aminopyrazole (13.7 g) in tetrahydrofuran (140 ml) ) Was added. The mixture was stirred at rt for 18 h and evaporated in vacuo. The residue was purified by column chromatography on silica gel eluting with ethyl acetate / hexanes (2: 3). The eluate was concentrated in vacuo to afford 4- (3-t-butoxycarbonylaminopropyl) -1- (2-triphenylmethyloxyethyl) -5-t-butoxycarbonylaminopyrazole (2.45 g) It was.

Example 14

Benzylhydryl 7β-[(Z) -2- (5-t-butoxycarbonylamino-1,2,4-thiadiazol-3-yl)-in N, N-dimethylformamide (2 ml)- To a suspension of 2- (1-t-butoxycarbonyl-1-methylethoxyimino) acetamido] -3-chloromethyl-3-cepem-4-carboxylate (1 g) and sodium iodide (182 mg) 4- (3-t-butoxycarbonylaminopropyl) -1- (2-triphenylmethyloxyethyl) -5-t-butoxycarbonylaminopyrazole (1.52 g) was added and the mixture was allowed to stand at room temperature for 24 Stir for hours. The reaction mixture was added to a mixture of ethyl acetate and water. The organic layer was separated and dried over magnesium sulfate. Magnesium sulfate was filtered and the filtrate was evaporated to about 20 ml under reduced pressure. The concentrate was poured into diisopropyl ether (150 ml) and the resulting precipitate was collected by filtration and dried in vacuo. To the resulting solid solution in methylene chloride (3 ml) was added anisole (1 ml) and trifluoroacetic acid (2 ml). The resulting solution was stirred at room temperature for 3 hours and poured into diisopropyl ether. The resulting precipitate was collected by filtration and dried in vacuo. The resulting powder was dissolved in phosphate buffer (pH 7) and adjusted to about pH 6 using saturated aqueous sodium hydrogen carbonate solution. The solution containing the desired compound was purified by preparative HPLC using an ODS column. The eluate containing the desired product was concentrated in vacuo. The concentrate was adjusted to about pH 2 with concentrated hydrochloric acid and eluted with 20% aqueous 2-propanol and chromatographed on Diaion ^R HP-20 (Mitsubishi Chemical Corporation). The eluate was evaporated in vacuo and lyophilized to yield 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxy. Mino) acetamido] -3- [3-amino-4- (3-aminopropyl) -2- (2-hydroxyethyl) -1-pyrazolo] methyl-3-cepem-4-carboxylate ( 61 mg) was obtained as an intangible solid.

Preparation Example 42

To a solution of 5-aminopyrazole-carbonitrile (102 g) and triethylamine (316 ml) in 1,4-dioxane (2000 ml) was added 1,3-dibromopropane (115 ml) and the mixture was 95 Stir at 5 ° C. for 5 days. Insoluble material was filtered off and the filtrate was concentrated in vacuo. 10% citric acid aqueous solution was added to the residue, and the mixture was extracted with chloroform. The extract was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with 2-propanol and dried in vacuo to give 4,5,6,7-tetrahydropyrazolo [1,5-a] pyridine-3-carbonitrile (8.0 g) as a solid.

Preparation Example 43

To a solution of 4,5,6,7-tetrahydropyrazolo [1,5-a] pyridine-3-carbonitrile (6.9 g) in pyridine (70 ml) was added triphenylmethyl chloride (15.6 g) and the mixture was Stir at 65 ° C. for 4 hours. After evaporation of the solvent in vacuo, the residue was dissolved in chloroform and the solution was washed with 10% aqueous citric acid solution. The organic layer was separated and the aqueous layer was extracted with chloroform. The organic layers were mixed, dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with diisopropyl ether and dried in vacuo to afford 4-triphenylmethyl-4,5,6,7-tetrahydropyrazolo [1,5-a] pyridine-3-carbonitrile (16.2 g). Obtained as a solid.

Preparation Example 44

To a solution of lithium aluminum hydride (2.7 g) in tetrahydrofuran (150 ml) under ice cooling, 4-triphenylmethyl-4,5,6,7-tetrahydropyrazolo [1,5-a] pyridine-3-carbo Nitrile (8.5 g) was added. The mixture was stirred at 65 ° C. for 16 h. After cooling in an ice bath, potassium fluoride (16.5 g) and water (5 ml) were added to the reaction mixture. Insoluble matter was removed by filtration. The filtrate was concentrated in vacuo to give 1- (4-triphenylmethyl-4,5,6,7-tetrahydropyrazolo [1,5-a] pyridin-3-yl) methylamine (8.4 g) as a solid. It was.

Preparation Example 45

Suspension of 1- (4-triphenylmethyl-4,5,6,7-tetrahydropyrazolo [1,5-a] pyridin-3-yl) methylamine (8.4 g) in ethyl formate (30 ml) Stir at 50 ° C. for 16 h. After evaporation of the solvent in vacuo, the residue was triturated with ethyl and dried in vacuo to give N-[(4,5,6,7-tetrahydropyrazolo [1,5-a] pyridin-3-yl) methyl] Formamide (3.0 g) was obtained as a rule solid.

Preparation Example 46

The silylated N-[(4,5,6,7-tetrahydropyrazolo [1,5-a] pyridin-3-yl) methyl] formamide solution in methylene chloride was diluted with methylene chloride (10 N-[(4,5,6,7-tetrahydropyrazolo [1,5-a] pyridin-3-yl) methyl] formamide (1.05 g), trimethylsilyl iodide (0.83 ml) in ml) And a mixture of diisopropylethylamine (1.02 ml) was prepared by stirring. Sodium iodide (291 mg) was added to a solution of benzylhydryl 7β-t-butoxycarbonylamino-3-chloromethyl-3-cepem-4-carboxylate (1.0 g) in N, N-dimethylformamide (1 ml). Was added and the mixture was stirred at rt for 30 min. To the resulting solution was added the above prepared silylated N-[(4,5,6,7-tetrahydropyrazolo [1,5-a] pyridin-3-yl) methyl] formamide solution and the reaction mixture was brought to room temperature. Stirred for 3.5 h. The reaction mixture was added to ethyl acetate and 2 mol / l aqueous calcium hydrogen sulfate solution. The aqueous layer was separated and the organic layer was washed with brine, dried over sodium sulfate and filtered. The filtrate was concentrated to about 20 ml in vacuo. The concentrate was poured into diisopropyl ether (300 ml) and the resulting precipitate was collected by filtration and dried in vacuo. To a suspension of solids produced in methylene chloride (3.0 ml) was added anisole (1.0 ml) and trifluoroacetic acid (2.0 ml). The resulting solution was stirred at rt for 3 h and poured into diisopropyl ether (300 ml). The resulting precipitate was collected by filtration and dried in vacuo to afford crude 7β-amino-3- [3- (formamidomethyl) -4,5,6,7-tetrahydro-1-pyrazolo [1,5-a] Pyridinio] methyl-3-cepem-4-carboxylate bistrifluoroacetic acid salt (810 mg) was obtained. This product was used in the next step without further purification.

Example 15

Crude 7β-amino-3- [3- (formamidomethyl) -4,5,6,7-tetra in N, N-dimethylformamide (16 ml) and tetrahydrofuran (16 ml) mixed solvent at room temperature N-trimethylsilylacetamide (2.94 g) was added to a solution of hydro-1-pyrazolo [1,5-a] pyridinio] methyl-3-cepem-4-carboxylate bistrifluoroacetic acid salt (800 mg). It was. The mixture was stirred at rt for 15 min. To the reaction mixture under ice cooling (Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-t-butoxycarbonyl-1-methylethoxyimino Acetyl chloride hydrochloride salt (420 mg) was added. The mixture was stirred for 2 hours under ice cooling. The reaction mixture was poured into ethyl acetate (250 ml) and the mixture stirred for 30 minutes. The resulting precipitate was collected by filtration, washed successively with ethyl acetate and diisopropyl ether, and dried in vacuo to give a solid (610 mg).

Anisole (0.6 ml) and trifluoroacetic acid (1.2 ml) were added to the resulting suspension of solid in methylene chloride (1.8 ml) under ice cooling. The resulting solution was stirred at rt for 3 h and poured into diisopropyl ether (150 ml). The resulting precipitate was collected by filtration and dried in vacuo to afford the crude product, which was purified by preparative HPLC using an ODS column. The eluate containing the desired product was concentrated to about 30 ml in vacuo. The concentrate was adjusted to ca. pH 3 with concentrated hydrochloric acid and chromatographed on Daion ^R HP-20 (Mitsubishi Chemical Corporation) eluting with 30% aqueous 2-propanol. The eluate was concentrated to about 30 ml in vacuo and lyophilized to afford 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1- Methylethoxyimino) acetamido] -3- [3- (formamidomethyl) -4,5,6,7-tetrahydro-1-pyrazolo [1,5-a] pyridinio] methyl-3 Cefem-4-carboxylate (130 mg) was obtained as an amorphous solid.

Example 16

7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methyl in 10% aqueous hydrochloric acid (4.36 ml) at room temperature Ethoxyimino) acetamido] -3- [3- (formamidomethyl) -4,5,6,7-tetrahydro-1-pyrazolo [1,5-a] pyridinio] methyl-3- The cefe-4-carboxylate (130 mg) solution was stirred for 6 hours. The reaction mixture was purified by preparative HPLC using an ODS column. The eluate containing the desired product was concentrated to about 30 ml in vacuo. The concentrate was adjusted to ca. pH 3 with concentrated hydrochloric acid and chromatographed on Daion ^R HP-20 (Mitsubishi Chemical Corporation) eluting with 30% aqueous 2-propanol. The eluate was concentrated to about 30 ml in vacuo and lyophilized to afford 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1- Methylethoxyimino) acetamido] -3- (3-aminomethyl-4,5,6,7-tetrahydro-1-pyrazolo [1,5-a] pyridinio) methyl-3-cepem-4 -Carboxylate (48 mg) was obtained as an intangible solid.

Preparation 47

Phosphorus oxychloride (123 ml) was added dropwise to N, N-dimethylformamide (307 ml) under ice cooling. To the mixture was added 4,5,6,7-tetrahydropyrazolo [1,5-a] pyridine-4-carbaaldehyde (100 g) solution in N, N-dimethylformamide (200 ml) under ice cooling. The reaction mixture was stirred at 80 ° C. for 2 hours. After cooling, water was added to the reaction mixture and the mixture was neutralized with sodium carbonate. The mixture was extracted with chloroform. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with hexane and dried in vacuo to give 4,5,6,7-tetrahydropyrazolo [1,5-a] pyridine-3-carbaaldehyde (65 g) as a solid.

Preparation Example 48

Triethyl phosphonoacetate (81.0 g) was added dropwise to a sodium hydride suspension (60% dispersion in mineral oil, 14.4 g) in tetrahydrofuran (300 ml) under ice cooling. The mixture was stirred for 1 h under ice cooling. 4,5,6,7-tetrahydropyrazolo [1,5-a] pyridine-3-carbaaldehyde (45.5 g) was added to the reaction mixture, and the mixture was stirred at room temperature for 17 hours. After evaporation of the solvent in vacuo, the residue was dissolved in chloroform and the solution was washed with water. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with diethyl ether and dried in vacuo to afford ethyl 3- (4,5,6,7-tetrahydropyrazolo [1,5-a] pyridin-3-yl) acrylate (49.2 g) as a solid. Obtained as

Preparation 49

Solution of ethyl 3- (4,5,6,7-tetrahydropyrazolo [1,5-a] pyridin-3-yl) acrylate (45 g) in a mixture of ethanol (800 ml) and tetrahydrofuran (400 ml) Was treated with 10% palladium carbon (2.2 g) at room temperature under hydrogen atmosphere at room temperature. After filtration of the catalyst the filtrate was concentrated in vacuo to give ethyl 3- (4,5,6,7-tetrahydropyrazolo [1,5-a] pyridin-3-yl) propionate (50 g) as an oil. It was.

Preparation 50

A mixture of ethyl 3- (4,5,6,7-tetrahydropyrazolo [1,5-a] pyridin-3-yl) propionate (20 g) and 1 mol / l aqueous sodium hydroxide solution (100 ml) was stirred at room temperature. Stirred for 16 h. The reaction mixture was neutralized with concentrated hydrochloric acid and the mixture was concentrated in vacuo. The residue was dissolved in methanol and insoluble sodium chloride was removed by filtration. The filtrate was concentrated in vacuo to give as crude 3- (4,5,6,7-tetrahydropyrazolo [1,5-a] pyridin-3-yl) propionic acid oil. This product was used in the next step without further purification.

Acetic anhydride (60 ml) was added to formic acid (50 ml) and the mixture was stirred at 40 ° C. for 2 hours. The resulting mixture was added to the oily product [3- (4,5,6,7-tetrahydropyrazolo [1,5-a] pyridin-3-yl) propionic acid] and the mixture was stirred at room temperature for 15 hours. . After evaporation of the solvent in vacuo, water was added to the residue. The mixture was neutralized with saturated aqueous sodium hydrogen carbonate and concentrated in vacuo. The residue was dissolved in methanol and insoluble material was removed by filtration. The filtrate was concentrated in vacuo to give as 3- (4-formyl-4,5,6,7-tetrahydropyrazolo [1,5-a] pyridin-3-yl) propionic acid (23 g) oil.

Preparation Example 51

3- (4-formyl-4,5,6,7-tetrahydropyrazolo [1,5-a] pyridin-3-yl) propionic acid (18.0 g), triethylamine (9.8 g) under ice cooling and To the mixture of tetrahydrofuran (300 ml) was added dropwise ethyl chloroformate (9.6 g). The mixture was stirred for 30 minutes under ice cooling. To the reaction mixture was added a solution of sodium azide (6.68 g) in water (100 ml) and the mixture was stirred at room temperature for 30 minutes. Methylene chloride was added to the reaction mixture and the aqueous layer was separated. The organic layer was washed with water, dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. Methanol (100 ml) was added to the oily residue and the mixture was stirred at reflux for 1.5 h. The reaction mixture was concentrated in vacuo to afford crude methyl N- [2- (4-formyl-4,5,6,7-tetrahydropyrazolo [1,5-a] pyridin-3-yl) ethyl] carbamate Obtained as an oil. This product was used in the next step without further purification.

Crude product [methyl N- [2- (4-formyl-4,5,6,7-tetrahydropyrazolo [1,5-a] pyridin-3-yl) ethyl] carbamate] and concentrated hydrochloric acid (60 ml) The mixture was stirred at reflux for 16 h. Water was added to the reaction mixture, and the mixture was adjusted to pH 5 using aqueous sodium hydroxide solution. The solution was washed with methylene chloride and the aqueous layer was adjusted to pH 8 using saturated aqueous sodium hydrogen carbonate solution. Di-t-butyl dicarbonate (10.0 g) was added to the solution, and the mixture was stirred at room temperature for 1 hour. The reaction mixture was extracted with methylene chloride. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. To the oily residue was added a solution of methanol (50 ml) and 4 mol / l hydrogen chloride in dioxane (50 ml) and the mixture was stirred at reflux for 30 minutes. After evaporation of the solvent in vacuo, the residue was triturated with 2-propanol and dried in vacuo to give 2- (4,5,6,7-tetrahydropyrazolo [1,5-a] pyridin-3-yl) ethyl Amine dihydrochloride (5.5 g) was obtained as a solid.

Preparation Example 52

2- (4,5,6,7-tetrahydropyrazolo [1,5-a] pyridin-3-yl) ethylamine dihydrochloride (2.36 g) and ethyl formate (40 g) in methanol (20 ml) To the solution was added a 28% sodium methoxide solution in methanol (4.0 g) and the mixture was stirred at reflux for 16 h. Insoluble material was filtered off using celite and the filtrate was concentrated in vacuo. The residue was triturated with acetonitrile and dried in vacuo to give N- [2- (4,5,6,7-tetrahydropyrazolo [1,5-a] pyridin-3-yl) ethyl] formamide (1.2 g ) Was obtained as a solid.

Preparation Example 53

7β-amino-3- [3- (2-formamidoethyl)-(4,5,6,7-tetrahydro-1-pyrazolo [1,5-a] pyridinio] methyl-3-cepem- 4-carboxylate bistrifluoroacetic acid salt

The title compound was prepared in the same manner as for 46, to prepare benzylhydryl 7β-t-butoxycarbonylamino-3-chloromethyl-3-cepem-4-carboxylate and N- [2- (4,5,6, Obtained from 7-tetrahydropyrazolo [1,5-a] pyridin-3-yl) ethyl] formamide.

Example 17

7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxyimino) acetamido] -3- [3- (2-formamidoethyl) -4,5,6,7-tetrahydro-1-pyrazolo [1,5-a] pyridinio] methyl-3-cepem-4-carboxylate

In the same manner as in Example 15, the title compound was prepared by 7β-amino-3- [3- (2-formamidoethyl)-(4,5,6,7-tetrahydro-1-pyrazolo [1,5- a] pyridinio] methyl-3-cepem-4-carboxylate bistrifluoroacetic acid salt.

Example 18

7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxyimino) acetamido] -3- [3- (2-aminoethyl) -4,5,6,7-tetrahydro-1-pyrazolo [1,5-a] pyridinio] methyl-3-cepem-4-carboxylate

In the same manner as in Example 16, the title compound was prepared by 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methyl. Ethoxyimino) acetamido] -3- [3- (2-formamidoethyl) -4,5,6,7-tetrahydro-1-pyrazolo [1,5-a] pyridinio] methyl- Obtained from 3-cefe-4-carboxylate.

Preparation Example 54

A solution of ethyl 3- (4,5,6,7-tetrahydropyrazolo [1,5-a] pyridin-3-yl) propionate (30 g) in methanol ammonium saturated solution (700 ml) was quenched at 110 ° C. Stir in autoclave for days. After evaporating the solvent in vacuo, methylene chloride was added to the residue. The resulting solid was collected by filtration and dissolved in methanol. Activated carbon was added to the solution, and the mixture was filtered. The filtrate was concentrated in vacuo. The residue was triturated with acetonitrile and dried in vacuo to afford 3- (4,5,6,7-tetrahydropyrazolo [1,5-a] pyridin-3-yl) propionamide (11.3 g) as a solid. It was.

Preparation Example 55

In a suspension of lithium aluminum hydride (7.6 g) in tetrahydrofuran (300 ml) at room temperature, 3- (4,5,6,7-tetrahydropyrazolo [1,5-a] pyridin-3-yl) propionamide ( 10 g) was added. The mixture was stirred at reflux for 2 days. After cooling in an ice bath, potassium fluoride (34 g) and water (10 ml) were added to the reaction mixture. Insoluble matter was removed by filtration. To the filtrate was added 10% aqueous sodium carbonate solution (100 ml) and di-t-butyl dicarbonate (20.0 g). The mixture was stirred at rt for 1 h. After evaporation of the solvent in vacuo, the residue was dissolved in chloroform. The solution was washed with water and dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. To the oily residue was added a solution of methanol (100 ml) and 4 mol / l hydrogen chloride in dioxane (50 ml) and the mixture was stirred at reflux for 30 minutes. The reaction mixture was concentrated in vacuo to give as crude 3- (4,5,6,7-tetrahydropyrazolo [1,5-a] pyridin-3-yl) propylamine (7.0 g) oil. This product was used in the next step without further purification.

To a solution of the crude product [3- (4,5,6,7-tetrahydropyrazolo [1,5-a] pyridin-3-yl) propylamine] (7.0 g) in methanol (60 ml) methanol (12 g) A 28% sodium methoxide solution was added followed by ethyl formate (120 g). The mixture was stirred at reflux for 20 h. The mixture was filtered through celide and the filtrate was concentrated in vacuo. The oily residue was purified by column chromatography on silica gel eluting with chloroform / methanol to give N- [3- (4,5,6,7-tetrahydropyrazolo [1,5-a] pyridin-3-yl). Propyl] formamide (3.1 g) was obtained as an oil.

Preparation Example 56

7β-amino-3- [3- (3-formamidopropyl)-(4,5,6,7-tetrahydro-1-pyrazolo [1,5-a] pyridinio] methyl-3-cepem- 4-carboxylate bistrifluoroacetic acid salt

In the same manner as in Preparation 46, the title compound was prepared with benzylhydryl 7β-t-butoxycarbonylamino-3-chloromethyl-3-cepem-4-carboxylate and N- [3- (4,5,6, Obtained from 7-tetrahydropyrazolo [1,5-a] pyridin-3-yl) propyl] formamide.

Example 19

7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxyimino) acetamido] -3- [3- (3-formamidopropyl) -4,5,6,7-tetrahydro-1-pyrazolo [1,5-a] pyridinio] methyl-3-cepem-4-carboxylate

In the same manner as in Example 15, the title compound was prepared by 7β-amino-3- [3- (3-formamidopropyl)-(4,5,6,7-tetrahydro-1-pyrazolo [1,5- a] pyridinio] methyl-3-cepem-4-carboxylate bistrifluoroacetic acid salt.

Example 20

7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxyimino) acetamido] -3- [3- (3-aminopropyl) -4,5,6,7-tetrahydro-1-pyrazolo [1,5-a] pyridinio] methyl-3-cepem-4-carboxylate

In the same manner as in Example 16, the title compound was prepared by 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methyl. Ethoxyimino) acetamido] -3- [3- (3-formamidopropyl) -4,5,6,7-tetrahydro-1-pyrazolo [1,5-a] pyridinio] methyl- Obtained from 3-cefe-4-carboxylate.

Preparation Example 57

N, N-dimethylformamide (339 ml) was added to phospho trichloride (136 ml) under ice cooling, and the mixture was stirred for 30 minutes under ice cooling. To the mixture was added a solution of 2,3-dihydro-1H-imidazo [1,2-b] pyrazole-1-carbaaldehyde (100 g) in N, N-dimethylformamide (200 ml) and the mixture was heated to 80 ° C. Stirred for 1.5 h. The reaction mixture was carefully added to a mixture of ethyl acetate and water under ice cooling, and the mixture was adjusted to pH 6. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with diisopropyl ether and dried in vacuo to afford 2,3-dihydro-1H-imidazo [1,2-b] pyrazole-1,7-dicarbaaldehyde (102 g).

Preparation 58

A solution of 2,3-dihydro-1H-imidazo [1,2-b] pyrazole-1,7-dicarbaaldehyde (1 g) and ammonium acetate (653 mg) in nitromethane (20 ml) was added for 4 hours. It was refluxed. The reaction mixture was evaporated under reduced pressure and extracted with dichloromethane. The extract was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo to give 7-[(E) -2-nitroethenyl] -2,3-dihydro-1H-imidazo [1,2-b] pyrazole-1 -Carbaaldehyde (1.00 g) was obtained.

Preparation Example 59

To a suspension of lithium aluminum hydride (465 mg) in tetrahydrofuran (10 ml) under ice cooling 7-[(E) -2-nitroethenyl] -2,3-dihydro-1H-imidazo [1,2- b] pyrazole-1-carbaaldehyde (300 mg) was added. The mixture was refluxed for 1.5 h. After cooling in an ice bath, sodium fluoride (2.06 g) and water (0.882 ml) were added to the reaction mixture. Insoluble matter was removed by filtration. The filtrate was concentrated in vacuo to afford 2- (2,3-dihydro-1H-imidazo [1,2-b] pyrazol-7-yl) ethylamine (165 mg). This product was used in the next step without further purification.

APC-MASS: m / z = 153 (M + H)

Preparation Example 60

A suspension of 2- (2,3-dihydro-1H-imidazo [1,2-b] pyrazol-7-yl) ethylamine (155 mg) in ethyl formate (3 ml) was refluxed for 4.5 hours. After evaporation of the solvent in vacuo, the residue was triturated with ethyl and dried in vacuo to afford 2- (2,3-dihydro-1H-imidazo [1,2-b] pyrazol-7-yl) ethylformamide (165 mg) obtained as a solid.

Preparation Example 61

Triphenylmethyl chloride in a solution of 2- (2,3-dihydro-1H-imidazo [1,2-b] pyrazol-7-yl) ethylformamide (1.67 g) in pyridine (15 ml) at room temperature 3.1 g) was added. The mixture was stirred at 60 ° C. for 5.5 h. The reaction mixture was evaporated under reduced pressure and extracted with ethyl acetate. The extract was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with ethyl acetate to give 2- (1-trityl-2,3-dihydro-1H-imidazo [1,2-b] pyrazol-7-yl) Ethylformamide (610 mg) was obtained.

Example 21

Benzylhydryl 7β-[(Z) -2- (5-t-butoxycarbonylamino-1,2,4-thiadiazol-3-yl)-in N, N-dimethylformamide (1 ml)- Mixture of 2- (1-t-butoxycarbonyl-1-methylethoxyimino) acetamido] -3-chloromethyl-3-cepem-4-carboxylate (0.5 g) and sodium iodide (99.6 mg) To a suspension of 2- (1-trityl-2,3-dihydro-1H-imidazo [1,2-b] pyrazol-7-yl) ethylformamide (587 mg) was added and the mixture was stirred at room temperature. Stir for 24 hours. The reaction mixture was added to a mixture of ethyl acetate and water. The organic layer was separated and dried over magnesium sulfate. Magnesium sulfate was filtered and the filtrate was evaporated to about 20 ml under reduced pressure. The concentrate was poured into diisopropyl ether (80 ml) and the resulting precipitate was collected by filtration and dried in vacuo. To the resulting solid solution in methylene chloride (3 ml) was added anisole (1 ml) and trifluoroacetic acid (2 ml). The resulting solution was stirred at room temperature for 3 hours and poured into diisopropyl ether. The resulting precipitate was collected by filtration and dried in vacuo. The resulting powder was dissolved in phosphate buffer (pH 7) and adjusted to about pH 6 using saturated aqueous sodium hydrogen carbonate solution. The solution containing the desired compound was purified by preparative HPLC using an ODS column. The eluate containing the desired product was concentrated in vacuo. The concentrate was adjusted to about pH 2 with concentrated hydrochloric acid and chromatographed on Daion ^R HP-20 (Mitsubishi Chemical Corporation) eluting with 20% aqueous 2-propanol. The eluate was concentrated to about 30 ml in vacuo and lyophilized to afford 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1- Methylethoxyimino) acetamido] -3- [2,3-dihydro-7- (2-formamidoethyl) -5- (1H-imidazo [1,2-b] pyrazolo)] Methyl-3-cepem-4-carboxylate (154 mg) was obtained as an amorphous solid.

Example 22

7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxyimino in methanol (1.5 ml) at room temperature ) Acetamido] -3- [2,3-dihydro-7- (2-formamidoethyl) -5- (1H-imidazo [1,2-b] pyrazolo)] methyl-3- To the solution of cefem-4-carboxylate (147 mg) was added concentrated hydrochloric acid (0.15 ml). The mixture was stirred at rt for 4 h and poured into ethyl acetate. The resulting precipitate was collected by filtration and dried in vacuo. The resulting powder was dissolved in phosphate buffer (pH 7) and adjusted to about pH 6 using saturated aqueous sodium hydrogen carbonate solution. The solution containing the desired compound was purified by preparative HPLC using an ODS column. The eluate containing the desired product was concentrated in vacuo. The concentrate was adjusted to about pH 2 with concentrated hydrochloric acid and chromatographed on Daion ^R HP-20 (Mitsubishi Chemical Corporation) eluting with 20% aqueous 2-propanol. The eluate was concentrated to about 30 ml in vacuo and lyophilized to afford 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1- Methylethoxyimino) acetamido] -3- [2,3-dihydro-7- (2-aminoethyl) -5- (1H-imidazo [1,2-b] pyrazoloio)] methyl- 3-cepem-4-carboxylate (32.4 mg) was obtained.

Preparation Example 62

Triethyl phosphonoacetate (80 ml) was added dropwise to a suspension of sodium hydride (60% dispersion in mineral oil, 16 g) in tetrahydrofuran (600 ml) under ice cooling. The mixture was stirred for 1 h under ice cooling. 2,3-dihydro-1H-imidazo [1,2-b] pyrazole-1,7-dicarbaaldehyde (60 g) was added to the reaction mixture at room temperature, and the mixture was stirred at room temperature for 2.5 hours. To the reaction mixture was added saturated aqueous ammonium chloride solution and extracted with ethyl acetate. The extract was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with diisopropyl ether and dried in vacuo to afford ethyl (2E) -3- (1-formyl-2,3-dihydro-1H-imidazo [1,2-b] pyrazole-7- I) -2-propenoate (63.5 g) was obtained. This product was used in the next step without further purification.

Preparation Example 63

Ethyl (2E) -3- (1-formyl-2,3-dihydro-1H-imidazo [1,2-b] pyrazole-7 in a mixture of ethanol (200 ml) and tetrahydrofuran (500 ml) The -yl) -2-propenoate (40 g) solution was treated with 10% palladium carbon (10 g) for 4.5 h at room temperature under a hydrogen atmosphere. After filtration of the catalyst the filtrate was concentrated in vacuo to afford ethyl 3- (1-formyl-2,3-dihydro-1H-imidazo [1,2-b] pyrazol-7-yl) propanoate (40.1 g) was obtained. This product was used in the next step without further purification.

Preparation Example 64

Ethyl 3- (1-formyl-2,3-dihydro-1H-imidazo [1,2-b] pyrazol-7-yl) in a mixture of methanol (50 ml) and 28% aqueous ammonia (104 ml) The propanoate (10 g) solution was stirred for 41 hours at room temperature. After evaporating the solvent in vacuo, the residue was triturated with diisopropyl alcohol and dried in vacuo to afford 3- (2,3-dihydro-1H-imidazo [1,2-b] pyrazol-7-yl Propanamide (6.44 g) was obtained.

Preparation 65

Triphenylmethyl chloride (11.1 g) in a solution of 3- (2,3-dihydro-1H-imidazo [1,2-b] pyrazol-7-yl) propanamide (6 g) in pyridine (60 ml) at room temperature ) Was added. The mixture was stirred at 60 ° C. for 17 h. The reaction mixture was evaporated under reduced pressure and extracted with ethyl acetate. The extract was dried over anhydrous magnesium sulfate, filtered and evaporated in vacuo. The residue was triturated with diisopropyl ether and dried in vacuo to afford 3- (1-trityl-2,3-dihydro-1H-imidazo [1,2-b] pyrazol-7-yl) propanamide ( 11.2 g) was obtained.

Preparation 66

To a suspension of lithium aluminum hydride (1.8 g) in tetrahydrofuran (200 ml) at room temperature, 3- (1-trityl-2,3-dihydro-1H-imidazo [1,2-b] pyrazole-7- 1) propanamide (10 g) was added. The mixture was stirred at rt for 2.5 h. After cooling in an ice bath, sodium fluoride (7.95 g) and water (3.4 ml) were added to the reaction mixture. Insoluble matter was removed by filtration. Ethyl formate was added to the filtrate, and the mixture was refluxed for 24 hours. After evaporation of the solvent in vacuo, the residue was triturated with diisopropyl ether and dried in vacuo to afford 3- (1-trityl-2,3-dihydro-1H-imidazo [1,2-b] pyrazole -7-yl) propylformamide (6.62 g) was obtained.

Preparation Example 67

Benzylhydryl 7β-t-butoxycarbonylamino-3-chloromethyl-3-cepem-4-carboxylate (2.5 g) in a mixture of N, N-dimethylformamide (5 ml) and methylene chloride (8 ml) Sodium iodide (728 mg) was added to the solution, and the mixture was stirred at room temperature for 30 minutes. To the reaction mixture was added 3- (1-trityl-2,3-dihydro-1H-imidazo [1,2-b] pyrazol-7-yl) propylformamide (6.36 g). The whole mixture was stirred at rt for 26 h and poured into a mixture of ethyl acetate and water. The aqueous layer was separated and the organic layer was washed with brine, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated to about 10 ml in vacuo. The concentrate was poured into diisopropyl ether (300 ml) and the resulting precipitate was collected by filtration and dried in vacuo. Methylene chloride (21 ml) To the resulting solid solution was added anisole (7 ml) and trifluoroacetic acid (14 ml). The resulting solution was stirred at room temperature for 3 hours and poured into diisopropyl ether. The resulting precipitate was collected by filtration and dried in vacuo to afford 7β-amino-3- [2,3-dihydro-7- (3-formamidopropyl) -5- (1H-imidazo [1,2-b] Pyrazolio)] methyl-3-cepem-4-carboxylate bistrifluoroacetate (4.17 g) was obtained. This product was used in the next step without further purification.

Example 23

Crude 7β-amino-3- [2,3-dihydro-7- (3-formamidopropyl) in N, N-dimethylformamide (20 ml) and tetrahydrofuran (20 ml) mixed solvent under ice cooling -5- (1H-imidazo [1,2-b] pyrazolo)] methyl-3-cepem-4-carboxylate bistrifluoroacetate (4.17 g) and N-trimethylsilylacetamide (8.63 g) (Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-t-butoxycarbonyl-1-methylethoxyimino) acetyl chloride hydro Chloride salt (2.53 g) was added. The mixture was stirred for 2 hours under ice cooling. The reaction mixture was poured into ethyl acetate and the mixture was stirred for 30 minutes. The resulting precipitate was collected by filtration, washed successively with ethyl acetate and diisopropyl ether and dried in vacuo to give a solid (2.2 g).

Anisole (3 ml) and trifluoroacetic acid (6 ml) were added to the resulting solid suspension in methylene chloride (9 ml) under ice cooling. The resulting solution was stirred at room temperature for 3 hours and poured into diisopropyl ether. The resulting precipitate was collected by filtration and dried in vacuo. The resulting powder was dissolved in phosphate buffer (pH 7) and adjusted to about pH 6 using saturated aqueous sodium hydrogen carbonate solution. The solution containing the desired compound was purified by preparative HPLC using an ODS column. The eluate containing the desired product was concentrated in vacuo. The concentrate was chromatographed on Daion ^R HP-20 (Mitsubishi Chemical Corporation), eluted with 20% aqueous 2-propanol with concentrated hydrochloric acid. The eluate was evaporated in vacuo and lyophilized to yield 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxy. Mino) acetamido] -3- [2,3-dihydro-7- (3-formamidopropyl) -5- (1H-imidazo [1,2-b] pyrazolo)] methyl-3 -Cefem-4-carboxylate (391.2 mg) was obtained.

Example 24

7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxyimino in methanol (3.9 ml) at room temperature ) Acetamido] -3- [2,3-dihydro-7- (3-formamidopropyl) -5- (1H-imidazo [1,2-b] pyrazoloio)] methyl-3- To the solution of cefem-4-carboxylate (386 mg) was added concentrated hydrochloric acid (0.386 ml). The mixture was stirred at rt for 3 h. The reaction mixture was poured into a mixture of ethyl acetate (300 ml) and acetone (100 ml). The resulting precipitate was collected by filtration and dried in vacuo. The resulting powder was dissolved in phosphate buffer (pH 7) and adjusted to about pH 6 using saturated aqueous sodium hydrogen carbonate solution. The solution containing the desired compound was purified by preparative HPLC using an ODS column. The eluate containing the desired product was concentrated in vacuo. The concentrate was adjusted to about pH 2 with concentrated hydrochloric acid and chromatographed on Daion ^R HP-20 (Mitsubishi Chemical Corporation) eluting with 20% aqueous 2-propanol. The eluate was evaporated in vacuo and lyophilized to yield 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxy. Mino) acetamido] -3- [2,3-dihydro-7- (3-aminopropyl) -5- (1H-imidazo [1,2-b] pyrazoloio)] methyl-3-cepem 4-carboxylate (84 mg) was obtained.

Preparation Example 68

2- (5-amino-4-cyano-1H-pyrazol-1-yl) acetamide

The title compound was obtained in the same manner as in Preparation 64, ethyl 2- (5-amino-4-cyano-1H-pyrazol-1-yl) acetate.

Preparation Example 69

2- [4-cyano-5- (tritylamino) -1H-pyrazol-1-yl] acetamide

The title compound was obtained from 2- (5-amino-4-cyano-1H-pyrazol-1-yl) acetamide in the same manner as in Example 27.

Preparation 70

To a suspension of lithium aluminum hydride (3.73 g) in tetrahydrofuran (100 ml) at room temperature was added 2- [4-cyano-5- (tritylamino) -1H-pyrazol-1-yl] acetamide (5 g). Was added. The mixture was refluxed for 23 hours. After cooling in an ice bath, sodium fluoride (16.5 g) and water (7.1 ml) were added to the reaction mixture. Insoluble matter was removed by filtration. The filtrate was evaporated in vacuo and triturated with diisopropyl ether to give 1- (2-aminoethyl) -4- (aminomethyl) -5-tritylaminopyrazole (2.17 g).

Preparation Example 71

To a solution of 1- (2-aminoethyl) -4- (aminomethyl) -5-tritylaminopyrazole (1 g) in tetrahydrofuran (35 ml) was added di-t-butyl dicarbonate (1.65 g). . The mixture was stirred at rt for 16 h. The reaction mixture was added to a mixture of ethyl acetate and water. The aqueous layer was separated and the organic layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated in vacuo and ground with hexanes to afford 1- (2-t-butoxycarbonylaminoethyl) -4- (t-butoxycarbonylaminomethyl) -5-tritylaminopyrazole (1.27 g). Obtained.

Example 25

7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxyimino) acetamido] -3- (3-amino-4-aminomethyl-2- (2-aminoethyl) -1-pyrazolio] methyl-3-cepem-4-carboxylate

Benzylhydryl 7β-[(Z) -2- (5-t-butoxycarbonylamino-1,2,4-thiadiazol-3-yl) -2 in the same manner as in Example 10 -(1-t-butoxycarbonyl-1-methylethoxyimino) acetamido] -3-chloromethyl-3-cepem-4-carboxylate and 1- (2-t-butoxycarbonylaminoethyl ) -4- (t-butoxycarbonylaminomethyl) -5-tritylaminopyrazole.

Preparation Example 72

Under a nitrogen atmosphere, 10% palladium (125 g) and methanol (3.75 L) on activated carbon were placed in a 5.0 L three neck flask. 5-amino-4-cyano-1- (2-hydroxyethyl) -1H-pyrazole (250 g) was added to the mixture. The mixture was stirred at rt for 1 h under hydrogen atmosphere. To the reaction mixture was added a solution of 4N hydrogen chloride in methanol (1.65 L) and the mixture was stirred at room temperature under hydrogen atmosphere for 1.5 hours. Water (1.2 L) was added to the reaction mixture. The catalyst was filtered off, washed with 50% aqueous methanol and the filtrate was evaporated in vacuo. The residue was triturated with isopropanol / ethyl acetate (1: 1). The precipitate was collected by filtration on a glass filter, washed with ethyl and dried in vacuo to afford 5-amino-4-aminomethyl-1- (2-hydroxyethyl) -1H-pyrazole dihydrochloride (358 g) as a white solid. Obtained.

Preparation Example 73

5-amino-4-aminomethyl-1- (2-hydroxyethyl) -1H-pyrazole dihydrochloride (105 g) is dissolved in a mixture of tetrahydrofuran (1700 ml) and water (170 ml) and the solution of The pH was adjusted to 9 using 3N aqueous sodium hydroxide solution. To the solution was added dropwise a solution of di-t-butyl dicarbonate (173 g) in tetrahydrofuran (860 ml), and 3N aqueous sodium hydroxide solution was added to maintain the pH of the reaction mixture at 8.5-9.0. Stirring was continued for an additional 1 hour. The reaction mixture was extracted with ethyl acetate. The organic layer was washed with brine, dried over magnesium sulfate, filtered and evaporated in vacuo. The residue was triturated with diisopropyl ether. The precipitates were collected by filtration on a glass filter, washed with diisopropyl ether and dried in vacuo to afford 5-amino-4- (t-butoxycarbonylaminomethyl) -1- (2-hydroxyethyl) -1H-pyrazole (118g) was obtained.

Preparation Example 74

Triphenyl in a solution of 5-amino-4- (t-butoxycarbonylaminomethyl) -1- (2-hydroxyethyl) -1H-pyrazole (243 g) in N, N-dimethylformamide (1.82 L) Methyl chloride (581 g), triethylamine (452 ml) and N, N-dimethylaminopyridine (9.91 g) were added successively. The reaction mixture was stirred at 70 ° C. overnight. After cooling at room temperature, the reaction mixture was diluted with ethyl acetate, washed with water and brine (twice), dried over magnusulfate and filtered. The filtrate was evaporated in vacuo. The residue was purified by silica gel column chromatography to give 4- (t-butoxycarbonylaminomethyl) -5- (tritylamino) -1- [2- (trityloxy) ethyl] -1H-pyrazole. Obtained and crystallized from diisopropyl ether / hexane (1: 2). The resulting crystals were collected by filtration on a glass filter, washed with diisopropyl ether / hexane (1: 2) and dried in vacuo to afford the desired compound (480 g).

Example 26

Benzylhydryl 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-t in N, N-dimethylformamide (12 ml) To a solution of butoxycarbonyl-1-methylethoxyimino) acetamido] -3-chloromethyl-3-cepem-4-carboxylate (5.82 g) was added sodium iodide (1.44 g). After stirring for 1 hour at room temperature, 4- (t-butoxycarbonylaminomethyl) -5- (tritylamino) -1- [2- (trityloxy) ethyl] -1 H-pyrazole (14.8 g ) Was added to the mixture. Stirring was continued at 35 ° C. for 24 hours. The resulting mixture was poured into water and extracted with ethyl acetate. The organic layer was washed with brine, dried over magnesium sulfate, filtered and evaporated in vacuo. The residue was dissolved in ethyl acetate (50 ml) and added dropwise to diisopropyl ether (500 ml). The resulting precipitate was collected by filtration. The filter cake was washed with diisopropyl ether and dried over phosphorus pentoxide in vacuo. The solid (15.6 g) was dissolved in dichloromethane (47 ml) and anisole (16 ml) and trifluoroacetic acid (32 ml) were added successively to the solution. After stirring for 3 hours at room temperature the reaction mixture was poured into diisopropyl ether (500 ml). The precipitate was collected by filtration, washed with diisopropyl ether and dried over phosphorus pentoxide in vacuo. The crude product was dissolved in phosphate buffer (pH 7.0) and purified by preparative HPLC (eluent: pH 7.0 phosphate buffer and acetonitrile). The eluate was chromatographed on Daion ^R HP-20 (Mitsubishi Chemical Corporation) and lyophilized to yield 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxyimino) acetamido] -3- [3-amino-4-aminomethyl-2- (2-hydroxyethyl) -1-pyrazolo] methyl-3-cepem 4-carboxylate (940 mg) was obtained.

Example 27

7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxyimino) acetamido] -3- [3-amino-4-aminomethyl-2- (2-hydroxyethyl) -1-pyrazolio] methyl-3-cepem-4-carboxylate (5.0 g) was dissolved in water (100 ml) and 2.0 M sulfuric acid (4.0 ml) was added to the solution. The mixture was lyophilized to give crude 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxyimino) acet Amido] -3- [3-amino-4-aminomethyl-2- (2-hydroxyethyl) -1-pyrazolio] methyl-3-cepem-4-carboxylate hydrogensulfate (5.18 g) Obtained as a solid.

Amorphous solid (1.0 g) was dissolved in water (1.0 ml). Acetonitrile (5.0 ml) was added dropwise to the solution. After stirring for 2 hours at room temperature, white crystals were precipitated. The precipitated crystals were collected by filtration on a glass filter, washed with a small amount of water / acetonitrile (1: 5) and dried under reduced pressure to give 7β-[(Z) -2- (5-amino-1,2,4-thia Diazol-3-yl) -2- (1-carboxy-1-methylethoxyimino) acetamido] -3- [3-amino-4-aminomethyl-2- (2-hydroxyethyl) -1 -Pyrazolio] methyl-3-cepem-4-carboxylate hydrogensulfate (880 mg) was obtained as white crystals.

Preparation 75

To a mixture of lithium aluminum hydride (32.6 g) in tetrahydrofuran (1.3 L) under ice cooling (2E) -3- [1-methyl-5- (tritylamino) -1H-pyrazol-4-yl] -2 Propenonitrile (101.6 g) was added. The mixture was stirred for 4 hours. After cooling in an ice bath, sodium fluoride (100 g) and water (100 ml) were added to the reaction mixture. Insoluble matter was removed by filtration. The filtrate was concentrated in vacuo to afford 4- (3-aminopropyl) -1-methyl-5-tritylamino-1H-pyrazole (88.9 g).

Preparation Example 76

To a solution of 4- (3-aminopropyl) -1-methyl-5-tritylamino-1H-pyrazole (75 g) in tetrahydrofuran (700 ml) was added di-t-butyl dicarbonate (49.5 g). . The reaction mixture was stirred at room temperature for 3 hours. After evaporation of the solvent in vacuo, the residue was triturated with diisopropyl ether and dried in vacuo to give t-butyl 3- [1-methyl-5- (tritylamino) -1H-pyrazol-4-yl] propyl Carbamate (71.7 g) was obtained.

Example 28

Benzylhydryl 7β-[(Z) -2- (5-t-butoxycarbonylamino-1,2,4-thiadia in N, N-dimethylformamide (120 ml) and methylene chloride (80 ml) Zol-3-yl) -2- (1-t-butoxycarbonyl-1-methylethoxyimino) acetamido] -3-chloromethyl-3-cepem-4-carboxylate (40 g) and sodium iodide To a suspension of the mixture of (8 g) t-butyl 3- [1-methyl-5- (tritylamino) -1 H-pyrazol-4-yl] propylcarbamate (60 g) was added and the mixture was allowed to stand at room temperature for 16 hours. Was stirred. The reaction mixture was added to a mixture of ethyl acetate and water. The organic layer was separated and dried over magnesium sulfate. Magnesium sulfate was filtered and the filtrate was evaporated to about 20 ml under reduced pressure. The concentrate was poured into diisopropyl ether (150 ml) and the resulting precipitate was collected by filtration and dried in vacuo. The precipitate was purified by column chromatography on Daion ^R PA306 (Mitsubishi Chemical Corporation) TFA form (400 ml) eluting with tetrahydrofuran. The eluate was concentrated in vacuo. The residue was dissolved in methylene chloride (200 ml) and anisole (70 ml) and trifluoroacetic acid (140 ml) were added to the solution. The mixture was stirred at rt for 3 h and poured into diisopropyl ether. The resulting precipitate was collected by filtration and dried in vacuo. The resulting powder was dissolved in phosphate buffer (pH 7) and adjusted to about pH 6 using saturated aqueous sodium hydrogen carbonate solution. The solution containing the desired compound was purified by preparative HPLC using an ODS column. The eluate containing the desired product was concentrated in vacuo. The concentrate was adjusted to about pH 1 with concentrated hydrochloric acid and chromatographed on Daion ^R PA306 (Mitsubishi Chemical Corporation) eluting with 20% aqueous 2-propanol. The eluate was evaporated in vacuo and lyophilized to yield 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxy. Mino) acetamido] -3- [3-amino-4- (3-aminopropyl) -2-methyl-1-pyrazolo] methyl-3-cepem-4-carboxylate (6.1 g) was obtained. .

Example 29

Benzylhydryl 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-t in N, N-dimethylformamide (660 ml) -Butoxycarbonyl-1-methylethoxyimino) acetamido] -3-chloromethyl-3-cepem-4-carboxylate (330 g) and sodium iodide (74.8 g) in a mixture suspension of t-butyl 3- [ 1-methyl-5- (tritylamino) -1H-pyrazol-4-yl] propylcarbamate (282 g) was added and the mixture was stirred at rt for 16 h. The reaction mixture was added to a mixture of ethyl acetate and water. The organic layer was washed with water, brine and 10% aqueous sodium trifluoroacetate solution and then dried over magnesium sulfate. Magnesium sulfate was filtered and the filtrate was evaporated to about 3.3 kg under reduced pressure. The concentrate was poured into diisopropyl ether (33 L) and the resulting precipitate was collected by filtration and dried in vacuo.

Anisole (530 ml) and trifluoroacetic acid (1600 ml) were added to the resulting solid suspension in methylene chloride (1600 ml). The mixture was stirred at rt for 3 h and poured into diisopropyl ether. The resulting precipitate was collected by filtration and dried in vacuo. The resulting powder was dissolved in phosphate buffer (pH 7) and adjusted to about pH 6 using saturated aqueous sodium hydrogen carbonate solution. The solution containing the desired compound was purified by preparative HPLC using an ODS column. The eluate containing the desired product was concentrated in vacuo. The concentrate was adjusted to ca. pH 2 with concentrated hydrochloric acid and chromatographed on Daion ^R PA306 (Mitsubishi Chemical Corporation) eluting with 20% aqueous 2-propanol. The eluate was evaporated in vacuo and lyophilized to yield 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxy. Mino) acetamido] -3- [3-amino-4- (3-aminopropyl) -2-methyl-1-pyrazolo] methyl-3-cepem-4-carboxylate (78.4 g) was obtained. .

Example 30

7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxyimino) acet in water (180 ml) Amido] -3- [3-amino-4- (3-aminopropyl) -2-methyl-1-pyrazolo] methyl-3-cepem-4-carboxylate (22.5 g) solution using an ODS column And purified by preparative HPLC. The eluate containing the desired product was concentrated to about 1.5 L in vacuo. The concentrate was adjusted to about pH 1 with concentrated hydrochloric acid and chromatographed on Daion ^R PA306 (Mitsubishi Chemical Corporation) eluting with 30% aqueous 2-propanol. The eluate was concentrated to about 400 ml in vacuo. 2 mol / l sulfuric acid (16 ml) was added to the solution and the mixture was lyophilized to give sulfate (16 g) as an amorphous powder. The powder was dissolved in water (70 ml) and 2-propanol (80 ml) with stirring at room temperature. Stirring was continued for 4 hours at room temperature. The precipitated crystals were filtered to give 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (1-carboxy-1-methylethoxyimino) acet Amido] -3- [3-amino-4- (3-aminopropyl) -2-methyl-1-pyrazolio] methyl-3-cepem-4-carboxylate hydrogensulfate (13 g) was obtained.

X-Ray Powder Diffraction Analysis (Philips MPD 1880 X-Ray Integral Diffraction System)

2θ century

8.5180

14160

14.5500

15.3400

15.5300

16.5420

17.3600

19410

19.4260

20240

24.5240

25430

26400

28250

X-ray: Monochromatic CnKα radiation

Voltage: 40 KV / Current: 30 mA

This application is based on Application No. PR 4690, filed in Australia on May 1, 2001, and Application No. PR 5834, filed in Australia on June 20, 2001, the contents of which are incorporated by reference.

Claims (9)

A compound of formula [I], or a pharmaceutically acceptable salt thereof: Where A is lower alkylene or lower alkenylene; R ¹ is lower alkyl, hydroxy (lower) alkyl, protected hydroxy (lower) alkyl, amino (lower) alkyl or protected amino (lower) alkyl; R ² is hydrogen or an amino protecting group; R ¹ and R ² join together to form lower alkylene; R ³ and R ⁵ are independently amino or protected amino; R ⁴ is carboxy or protected carboxy. The compound of claim 1, wherein R ¹ is lower alkyl, hydroxy (lower) alkyl, aryl (lower) alkyloxy (lower) alkyl, amino (lower) alkyl or acylamino (lower) alkyl; R ² is hydrogen, aryl (lower) alkyl or acyl; R ¹ and R ² join together to form lower alkylene; R ³ and R ⁵ are independently amino or acylamino; R ⁴ is carboxy or esterified carboxy, or a pharmaceutically acceptable salt thereof. The compound of claim 2, wherein R ¹ is lower alkyl, hydroxy (lower) alkyl, aryl (lower) alkyloxy (lower) alkyl, amino (lower) alkyl, (lower) alkanoylamino (lower) alkyl, or (lower) Alkoxycarbonylamino (lower) alkyl; R ² is hydrogen, aryl (lower) alkyl, lower alkanoyl or lower alkoxycarbonyl; R ¹ and R ² join together to form lower alkylene; R ³ and R ⁵ are independently amino, lower alkanoylamino or lower alkoxycarbonylamino; R ⁴ is carboxy or lower alkoxycarbonyl, or a pharmaceutically acceptable salt thereof. The compound of claim 3, wherein R ¹ is lower alkyl, hydroxy (lower) alkyl or amino (lower) alkyl; R ² is hydrogen; R ¹ and R ² join together to form lower alkylene; R ³ and R ⁵ are amino; R ⁴ is carboxy, or a pharmaceutically acceptable salt thereof. (1) A compound of formula [II], or a reactive derivative in its amino group, or a salt thereof, is reacted with a compound of formula [III], or a reactive derivative in carboxy group thereof, or a salt thereof to To obtain a compound or a salt thereof, (2) removing the amino protective group to the compound of formula [Ia] or a salt thereof to obtain a compound of formula [Ib] or a salt thereof, or (3) reacting a compound of formula [VI] or a salt thereof with a compound of formula [VII], or a salt thereof, to obtain a compound of formula [VIII] or a salt thereof, A process for preparing a compound of formula [I] or a salt thereof, comprising performing a removal reaction of a carboxy protecting group on a compound of formula [VIII] or a salt thereof to obtain a compound of formula [I] or a salt thereof: Where A is lower alkylene or lower alkenylene; R ¹ is lower alkyl, hydroxy (lower) alkyl, protected hydroxy (lower) alkyl, amino (lower) alkyl or protected amino (lower) alkyl; R ² is hydrogen or an amino protecting group; R ¹ and R ² join together to form lower alkylene; R ³ and R ⁵ are independently amino or protected amino; R ⁴ is carboxy or protected carboxy; R ^3a is protected amino; R ⁶ is protected carboxy; Y is an exit group. A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof in admixture with a pharmaceutically acceptable carrier. A compound according to claim 1 or a pharmaceutically acceptable salt thereof for use as a medicament. A compound according to claim 1 or a pharmaceutically acceptable salt thereof for use as an antimicrobial agent. Use of the compound of claim 1 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of an infectious disease.